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		<div class="Basic-Text-Frame">
			<p class="Type-of-Article">&nbsp;</p>
			<p class="Type-of-Article"><span class="CharOverride-1">Research Article</span></p>
	</div>
		<div class="Basic-Text-Frame">
			<p class="title- ParaOverride-1">&nbsp;</p>
			<p class="title- ParaOverride-1">Sequence Analysis, 3-Dimensional Protein Modelling and Epitope Prediction of Oma87 Gene Orignating from Pasteurella multocida Isolate of Sheep</p>
		</div>
		<div class="Basic-Text-Frame">
			<p class="Authors">&nbsp;</p>
			<p class="Authors"><span class="CharOverride-3">Kumar Ragavendhar, Palanisamy Prabhakar, Ardhanary Thangavelu, John John Kirubaharan, Navamani Daniel Joy Chandran  </span> </p>
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		<div class="Basic-Text-Frame">
			<p class="Affiliations">Department of Veterinary Microbiology, Madras Veterinary College, Chennai – 600 007, India.</p>
		</div>
		<div>
			<p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-4">Abstract </span>| <span>Pasteurellosis, caused by </span><span class="CharOverride-5">Pasteurella multocida, </span><span>is an acute septicaemic disease characterized by high morbidity and mortality. Apart from cattle and buffaloes, it also results in severe economic losses in sheep and goat. Frequent reports of sheep pasteurellosis in India raised the need for specific and effective vaccination against sheep pasteurellosis. To identify antigenic determinants suitable for epitope based subunit vaccine, we have reported genome and protein sequence based characterization of outer membrane antigen 87 (</span><span class="CharOverride-5">Oma87</span><span>) of </span><span class="CharOverride-5">P. multocida </span><span>of</span><span class="CharOverride-5"> </span><span>sheep origin. </span><span class="CharOverride-5">Oma87</span><span> gene was amplified by PCR, purified and sequenced. Sequence analysis showed 99% homology with Pm70 and other </span><span class="CharOverride-5">P. multocida Oma87</span><span> genes available in the GenBank. Later, the 3-dimensional structure, T cell and B cell binding epitopes of the </span><span class="CharOverride-5">Oma87</span><span> protein were predicted using the </span><span class="CharOverride-5">O</span><span class="CharOverride-5">ma87</span><span> protein sequence. Our studies suggest that predicted </span><span class="CharOverride-6">T cell binding epitope (FYQYYSAGGIGSLRG) could be used either alone or together with predicted B cell epitope as a suitable epitope based subunit vaccine against </span><span class="CharOverride-5">P. multocida </span><span>infection.</span></p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-4">Keywords</span> | <span class="CharOverride-5">P. multocida Oma87</span> gene, Nucleotide sequence, Sequence analysis, Protein modelling,  T cell epitope.</p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
		</div>
		<div class="Basic-Text-Frame">
			<p class="Editor----Citation"><span class="CharOverride-7">Editor</span> | Tahir Yaqub, University of Veterinary and Animal Sciences, Lahore, Pakistan.</p>
			<p class="Editor----Citation"><span class="CharOverride-4">Received</span> | October 14, 2014; <span class="CharOverride-4">Revised</span> | October 30, 2014; <span class="CharOverride-4">Accepted</span> | October 31, 2014; <span class="CharOverride-4">Published</span> | November 15, 2014&#9;&#9;</p>
			<p class="Editor----Citation"><span class="CharOverride-4">*Correspondence</span> | Ardhanary Thangavelu, Madras Veterinary College, Chennai, India; <span class="CharOverride-4">Email:</span> thangavelu_a@hotmail.com</p>
			<p class="Editor----Citation"><span class="CharOverride-4">Citation</span> | Ragavendhar K, Prabhakar P,  Thangavelu A, Kirubaharan JJ, Chandran NDJ (2015). Sequence analysis, 3-dimensional protein modelling and epitope prediction of Oma87 gene orignating from Pasteurella multocida isolate of sheep. J. Inf. Mol. Biol. 3(1): 1-12.</p>
			<p class="Editor----Citation"><span class="CharOverride-4">DOI</span> | <a href="http://dx.doi.org/10.14737/journal.jimb/2015/3.1.1.12"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.jimb/2015/3.1.1.12</span></a>   </p>
			<p class="Editor----Citation"><span class="CharOverride-7">I</span><span class="CharOverride-4">SSN (Online)</span> |  2307-5465; <span class="CharOverride-4">ISSN (Print)</span>  | 2307-5716</p>
			<p class="Editor----Citation"><span class="CharOverride-7">Copyright </span>© 2015 Ragavendhar et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</p>
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			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
		  <p class="Heading-1--Introduction----">INTRODUCTION</p>
			<p class="Caps-on-First-Para ParaOverride-1">&nbsp;</p>
			<p class="Caps-on-First-Para ParaOverride-1"><span class="_idGenDropcap-1">T</span>he bacterium, <span class="CharOverride-5">Pasteurella multocida (P. multocida), </span>is an opportunistic pathogen responsible for fowl cholera in poultry, haemorrhagic septicaemia of buffalo, pneumonia of lambs and goats, respiratory atrophic rhinitis of swine and purulent rhinitis of rabbit (<a href="#Gyles-CL--Prescott-JF--Songer-J--Thoen-CO--2004-.-I"><span class="Hyperlink">Gyles et al., 2004</span></a>). Sheep pasteurellosis, caused by <span class="CharOverride-5">P. multocida </span>serotype A, is one of the common infectious diseases occurring in temperate and sub-tropical areas (<a href="#Chandrasekaran-S--1991-."><span class="Hyperlink">Chandrasekaran, 1991</span></a>). In India, efforts are being done to control this disease with the use of killed vaccine prepared from<span class="CharOverride-5"> P. multocida </span>P52 strain. Even though the same vaccine is used for sheep and goats, the outbreaks stills occurs in India as reported previously (<a href="#Prabhakar-TG--Prabhakar-P--Venkadabadi--2010-"><span class="Hyperlink">Prabhakar et al., 2010</span></a>; <a href="#Dar-LM--Darzi-MM--Mir-MS--Rashid-A--Abdullah-S--Hussain-SA--2012-."><span class="Hyperlink">Dar et al., 2012</span></a>; <a href="#Prabhakar-P--Thangavelu-A--Kirubaharan-JJ--Chandran-NDJ--2012-"><span class="Hyperlink">Prabhakar et al., 2012</span></a>). This urges the development of newer and more effective vaccines to be used for prophylaxis against pasteurellosis. In this regards, with the development of reverse vaccinology, vaccines could be developed with the aid of computational analysis of genome (<a href="#Amol-KM--Soni-R--Girdhar-A--Tiwari-A--2013-."><span class="Hyperlink">Amol et al., 2013</span></a>). </p>
		  <p class="Caps-on-First-Para ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Since immunization with lipopolysaccharides alone induced only partial protection, outer membrane proteins are thought to be the major antigens involved in inducing cross-protective response (<a href="#Adler-R--Adler-B--1991-"><span class="Hyperlink">Adler and Adler, 1991</span></a>). Several outer membrane proteins (OMP) are immunogens and the antibodies against them demonstrate a strong protective activity. Such antigens may be used as a component of a subunit vaccine which lacks the demerits of conventional vaccine (<a href="#Confer-AW--Suckow-MA--Montelongo-M--Dabo-SM--Miloscio-LJ--Gillespie-AJ--Meredith-GL--2001"><span class="Hyperlink">Confer et al., 2001</span></a>). Further, an 87-kDa outer membrane antigen coded by <span class="CharOverride-5">Oma87</span> gene has been reported in all 16 serotypes of <span class="CharOverride-5">P. multocida </span>(<a href="#Raffolo-CG--Adler-B--1996-."><span class="Hyperlink">Raffolo and Adler, 1996</span></a>). Added to this, <a href="#Singh-S--Singh-VP--Cheema-PS--Sandey-M--Ranjan-R--Kumargupta-S--2011-"><span class="Hyperlink">Singh et al. (2011)</span></a> have investigated outer membrane protein DNA for their potency to be used as successful vaccine against haemorrhagic septicaemia of cattle and buffaloes (<a href="#Singh-S--Singh-VP--Cheema-PS--Sandey-M--Ranjan-R--Kumargupta-S--2011-"><span class="Hyperlink">Singh et al.,</span><span class="Hyperlink"> </span><span class="Hyperlink">2011</span></a>). In this study, we report the sequencing and sequence analysis of gene encoding 87-kDa <span class="CharOverride-5">Oma87</span> gene of <span class="CharOverride-5">P. multocida </span>serotype ‘A’ isolate obtained from sheep as well as the generation of 3D protein structure of <span class="CharOverride-5">Oma87</span> protein with validation by VADAR and Ramachandran plot. We also employed reverse vaccinology based computational analysis for prediction of <span class="CharOverride-5">Oma87</span> protein specific B cell and T cell epitopes along with their docking studies to the MHC class-I and class-II alleles for the suitable vaccine development.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">MATERIALS AND METHOD</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-2--History-in-MM-"><span class="CharOverride-2">P. </span><span class="CharOverride-11">multocida</span><span class="CharOverride-2"> </span>Isolates</p>
			<p class="Body-Text ParaOverride-1">Thirteen <span class="CharOverride-5">P. multocida </span>sheep isolates were inoculated in BHI broth and incubated at 37<span class="CharOverride-12">o</span>C overnight. After incubation all the thirteen isolates were injected (0.2mL) via peritoneal route into thirteen Swiss albino mice and the mean death time was observed. Aspirated heart blood from dead mice was streaked on 10% sheep blood agar and incubated at 37<span class="CharOverride-12">o</span>C. Biochemical tests were carried out for identification. The biochemical tests included IMVIC, sugar fermentation test and catalase and oxidase test (<a href="#Office-International-des-epizooties--OIE---2008-."><span class="Hyperlink">OIE manual, 2008</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM- ParaOverride-1">DNA Extraction and <span class="CharOverride-2">P. </span><span class="CharOverride-11">multocida</span> Species Specific PCR (PM-PCR)</p>
			<p class="Body-Text ParaOverride-1">DNA was extracted from the culture by high salt method as described previously (<a href="#Kumar-S--Ramdass-P--2001-."><span class="Hyperlink">Kumar and Ramdass, 200</span></a><span class="Hyperlink">1</span>) and stored at –20<span class="CharOverride-12">o</span>C until used. Previously developed <span class="CharOverride-5">P. multocida </span>specific PCR assay (<a href="#Townsend-KM--Frost-AJ--Lee-CW--Papadimitriou-JM--Dawkins-HJS--1998-."><span class="Hyperlink">Townsend et al., 1998</span></a>) was used for the further confirmation of 13 <span class="CharOverride-5">P. multocida </span>isolates. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM- ParaOverride-1">PCR and Sequencing of <span class="CharOverride-5">Oma87</span> Gene of <span class="CharOverride-2">P. </span><span class="CharOverride-11">multocida</span></p>
			<p class="Body-Text ParaOverride-1">From the thirteen isolates, one <span class="CharOverride-5">P. multocida </span>sheep isolate was selected for amplification of <span class="CharOverride-5">Oma87</span> gene. Both forward 5´<span class="CharOverride-13">actcacaggataacattcacg</span> 3´ and Reverse 5´ <span class="CharOverride-13">ttttacataagttccaatcgc</span> 3´ primers were designed based on the <span class="CharOverride-5">Oma87</span> gene sequence (GenBank Accession No. U60439). The PCR reaction mixture contained 50ng of gDNA, 10 pmol of each primer and 1 units of hot start Taq polymerase. The thermal cycle protocol were as follows: Initial denaturation at 94<span class="CharOverride-12">o</span>C for 5 min, followed by 30 cycles, each cycle consisting of 3 steps- denaturation at 94<span class="CharOverride-12">o</span>C for 1 min, annealing at 49<span class="CharOverride-12">o</span>C for 1 min, Extension at 72<span class="CharOverride-12">o</span>C for 3 min. Final Extension was carried out at 72<span class="CharOverride-12">o</span>C for 10 min. The amplified (2.42kb) product was purified and the sequencing was carried out by primer walking. Sequence was submitted to NCBI GenBank (Accession number KJ576825).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM- ParaOverride-1">Sequence Analysis and Protein Modelling of <span class="CharOverride-5">Oma87</span> Protein</p>
			<p class="Body-Text ParaOverride-1">A global blast analysis of the selected sequence revealed the closest match of it.  About nine hits and representative sequences were selected from Blast results for the alignment. The sequences were aligned by clustal w and phylogenetic tree was constructed based on the clustal w alignment using MEGA 6 (<a href="#Tamura-K--Nei-M--Kumar-S--2004"><span class="Hyperlink">Tamura et al., 2004</span></a>; <a href="#Tamura-K--Stecher-G--Peterson-D--Filipski-A--Kumar-S--2013-."><span class="Hyperlink">Tamura et al., 2013</span></a>). This <span class="CharOverride-5">Oma87</span> protein sequence was subjected to protein structure modelling using SWISS–MODEL homology modeller (<a href="http://beta.swissmodel.expasy.org/workspace"><span class="Hyperlink">http://beta.swissmodel.expasy.org/workspace</span></a>) in an automated mode (<a href="#Benkert-P--Biasini-M--Schwede-T--2011-"><span class="Hyperlink">Benkert et al., 2011</span></a>). Validation was done by VADAR (<a href="#Willard-L--Ranjan-A--Zhang-H--Monzavi-H--Boyko-RF--Sykes-BD--David-S--2003"><span class="Hyperlink">Willard et al., 2003</span></a>) and Ramachandran plot using PROCHECK (<a href="#Ramachandran-GN--Ramakrishnan-C--Sasisekharan-V--1963-."><span class="Hyperlink">Ramachandran et al., 1963</span></a>; <a href="#Laskowski-RA--MacArthur-MW--Moss-DS--Thornton-JS--1993-"><span class="Hyperlink">Laskowski et al., 1993</span></a>). Physiochemical characterization was done by PROTPARAM software using ExPASyserver (<a href="#Ikai-AJ--1980-.-Ther"><span class="Hyperlink">Ikai et al., 1980</span></a>; <a href="#Kyte-J--Doolittle-RF--1982-"><span class="Hyperlink">Kyte and Doolittle, 1982</span></a>; <a href="#Guruprasad-K--Reddy-BVB--Pandit-MW--1990"><span class="Hyperlink">Guruprasad et al., 1990</span></a>; <a href="#Gasteiger-E--Hoogland-C--Gattiker-A--Duvaud-S--Wilkins-MR--Appel-RD--Bairoch-A--2005-"><span class="Hyperlink">Gasteiger et al., 2005</span></a>). The server VaxiJen (<a href="#Doytchinova-IA--Flower-DR--2007-"><span class="Hyperlink">Doytchinova et al., 2007</span></a>) was used to analyse the antigenicity of the <span class="CharOverride-5">Oma87</span> protein.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-2--History-in-MM- ParaOverride-1">T Cell and B Cell Epitope Prediction, 3D Structure Prediction of T Cell Epitopes, MHC Class II H2-IAB Allele, MHC Class I H2-Kda Allele and their Docking Studies <span class="CharOverride-5">Oma87</span> protein sequence was subjected to Mouse MHC class I allele and MHC class II allele binding T cell epitope prediction using T cell epitope prediction by IEDB-AR tools (tools.immuneepitope.org/mhc). The epitope having IC50 values within 250nm were selected and their antigen score was analysed using VaxiJen server. <span class="CharOverride-5">Oma87</span> protein sequence was subjected to B cell epitope prediction using ABC prediction server (<a href="#Saha-S--Raghava-GPS--2006-"><span class="Hyperlink">Saha and Raghava, 2006</span></a>). 3D structure of the selected epitope was predicted using PEP-fold 1.5 peptide structure prediction server at the RPBS Mobyle Portal (<a href="#Maupetit-J--Derreumaux-P--Tuffery-P--2009-."><span class="Hyperlink">Maupetit et al., 2009</span></a>; <a href="#Maupetit-J--Derreumaux-P--Tuffery-P--2010-.-A"><span class="Hyperlink">Maupetit et al., 2010</span></a>; <a href="#Thevenet-P--Shen-Y--Maupetit-J--Guyon-F--Derreumaux-P--Tuffery-P--2012"><span class="Hyperlink">Thevenet et al., 2012</span></a>). Best conformation given by the server was selected for further docking studies. Also the 3D structure of Mouse MHC class I H2-kDa allele and Mouse MHC class II H2-I-Ab allele were predicted using SWISS–MODEL homology modeller. Validation was done by Vadar and Ramachandran plot. For this purpose, protein sequence of both H2-kDa allele and H2-I-ab allele were obtained from NCBI gene bank database AAX98169.1 and NP_996988.2 respectively. The docking of the predicted <span class="CharOverride-5">Oma87</span> T cell epitopes to MHC I H2-kDa Protein and MHC II H2-I-ab protein were carried out using Patchdock server (<a href="#Schneidman-Duhovny-D--Inbar-Y--Nussinov-R--2005-."><span class="Hyperlink">Schneidman-Duhovny et al., 2005</span></a>) with the RMSD value 1.5A as recommended for protein small molecule docking and the solved structures were further refined using Firedock server (<a href="#Mashiach-E--Schneidman-Duhovny-D--Andrusier-N--2008-."><span class="Hyperlink">Mashiach et al., 2008</span></a>).</p>
			<p class="Heading-2--History-in-MM- ParaOverride-1">&nbsp;</p>
		  <p class="Heading-2--History-in-MM- ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">RESULTS AND DISCUSSION</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Pasteurellosis<span class="CharOverride-5"> </span>is an acute septicaemic disease characterized by high morbidity and mortality in cattle, sheep, goat and poultry resulting in economic losses. Pasteurellosis in sheep is capsular A serotype specific, which affects all age group of sheep leading to unthriftiness, reduction in body weight and death (<a href="#Prabhakar-TG--Prabhakar-P--Venkadabadi--2010-"><span class="Hyperlink">Prabhakar et al., 2010</span></a>). To develop an effective epitope based vaccine for sheep pasteurellosis, we focused on outer membrane protein of <span class="CharOverride-5">P. multocida </span>which is proved to be immunogenic.<span class="CharOverride-16"> </span>Recently, a study based on the immunoinformatics revealed 129 immunogens in <span class="CharOverride-5">P. multocida</span>. Further <span class="CharOverride-5">in vitro</span> gene expres sion studies resulted in twelve immunogenic determinants including <span class="CharOverride-5">Oma87</span>, Omp A, Omp16 and Plp B against <span class="CharOverride-5">P. multocida </span>infected chicken sera (<a href="#Al-Hasani-K--Boyce-J--McCarl-VP--Bottomley-S--Wilkie-I--Adler-B--2007-."><span class="Hyperlink">Al-Hasani et al., 2007</span></a>). Recombinant <span class="CharOverride-5">Oma87</span> protein developed against Fowl cholera causing <span class="CharOverride-5">P. multocida </span>A serotype showed effective protection in an experimental study with mice and rabbit (<a href="#Raffolo-CG--Adler-B--1996-."><span class="Hyperlink">Raffolo and Adler, 1996</span></a>). Immunization of mice with recombinant <span class="CharOverride-5">Oma87</span> protein developed against <span class="CharOverride-5">P. multocida </span>P52 strain B serotype showed increased antigen speciﬁc IgG titres in serum, whereas challenge studies showed 66.6 and 83.3% protection against <span class="CharOverride-5">P. multocida </span>B: 2 strain and <span class="CharOverride-5">P. multocida </span>A: 1 strain, respectively (<a href="#Kumar-A--Yogisharadhya-RR--Ramakrishnan-MA--Viswas-KN--Shivachandra-SB--2013-."><span class="Hyperlink">Kumar et al., 2013</span></a>). </p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			
          <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117032203.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117032203.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><a id="Figure-1"></a><span class="CharOverride-4">Figure 1: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117032203.png">Agarose gel electrophoresis analysis showing <span class="CharOverride-5">Oma87</span> gene (2.42 Kb) of <span class="CharOverride-5">P. multocida </span>sheep S02 isolate and 1Kb ladder</a></p>
       </div>
          <p>&nbsp;</p>
          <p><br>
          </p>
          <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Table-1"></a>Table 1: </span>History of the thirteen isolates of <span class="CharOverride-5">P. multocida </span>with origin, date of sample collection and their respective clinical symptoms</p>
			<table width="657" height="397" class="Table-Style-1 TableOverride-1" id="table-1">
				<colgroup>
					<col class="_idGenTableRowColumn-1" />
					<col class="_idGenTableRowColumn-2" />
					<col class="_idGenTableRowColumn-3" />
					<col class="_idGenTableRowColumn-3" />
					<col class="_idGenTableRowColumn-3" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-4">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">Isolates</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">MVC S01-S04</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">MVC S05-S07</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">MVC S08-S10</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">MVC S11-S13</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Location of outbreak/ sample collection</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Dharmapuri,</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">Tamil Nadu</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Dindigul,</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">Tamil   Nadu</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Kattupakkam,</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">Tamil Nadu</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Sunampet,</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">Tamil Nadu</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-6">
						<td>
							<p class="Body-Text ParaOverride-3"><span class="Title">Date of sample collection</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">24-06-08</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">25-06-08</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">12-07-08</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">24-12-08</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-7">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Species affected</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Ovine</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Ovine</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Ovine</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Ovine</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-8">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Age of animals</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1-3 years</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1-3 years</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1-3 years</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1-3 years</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-5">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Clinical symptoms</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Respiratory distress and nasal discharge</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Nasal discharge</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Respiratory distress and nasal discharge</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Nasal discharge</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-9">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">Mouse bio assay</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">(MDT)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">S01-15 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S02- 13 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S03- 15 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S04-17 hour</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">S05-15 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S06- 17 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S07- 16 hour</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">S08-15 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S09- 16 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S10- 16 hour</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">S11-20 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S12- 21 hour</span></p>
							<p class="Body-Text ParaOverride-1"><span class="Title">S13- 28 hour</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Normal" lang="en-US">&nbsp;</p>
			
          <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117030604.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117030604.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-2--P"></a>Figure 2: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117030604.png">Phylogenetic tree analysis for the <span class="CharOverride-5">Oma87</span> gene sequence of <span class="CharOverride-5">P. multocida </span>sheep S02 isolate with nine similar sequence obtained by BLAST hits using MEGA6</a>          </p>
          </div>
          <br>
			
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text">Based on the conventional microbiology techniques all the 13 isolates were identified as <span class="CharOverride-5">P. multocida </span>(<a href="#Office-International-des-epizooties--OIE---2008-."><span class="Hyperlink">OIE manual, 2008</span></a>) and further confirmed by<span class="CharOverride-5"> P. multocida </span>species specific PCR (PM–PCR) assay (<a href="#Townsend-KM--Frost-AJ--Lee-CW--Papadimitriou-JM--Dawkins-HJS--1998-."><span class="Hyperlink">Townsends </span></a><a href="#Townsend-KM--Frost-AJ--Lee-CW--Papadimitriou-JM--Dawkins-HJS--1998-."><span class="Hyperlink">et al, 1998</span></a>). In mouse biological assay ten <span class="CharOverride-5">P. multocida </span>isolates were found to be virulent with MDT between 12-18 hrs. Two isolates showed MDT between 19 and 24 hrs. One isolate showed MDT between 25 and 42 hrs (<a href="#Table-1"><span class="Hyperlink">Table 1</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Amplification of <span class="CharOverride-5">Oma87</span> gene from <span class="CharOverride-5">P. multocida</span> (SO2)<span class="CharOverride-5"> </span>isolate resulted in the amplified product of 2.42bp (<a href="#Figure-1"><span class="Hyperlink">Figure 1</span></a>). The amplified product was sequenced by primer walking. Only the complete 2373bp sequence of <span class="CharOverride-5">oma87</span> gene was submitted to NCBI GenBank under the accession number <span class="CharOverride-6">KJ576825 from the obtained product.</span> It showed 99% homology with <span class="CharOverride-5">P. multocida </span>subsp <span class="CharOverride-5">P. multocida</span> str. Pm70, <span class="CharOverride-5">P. multocida </span>omp87 gene for omp87 protein, strain P52, <span class="CharOverride-5">P. multocida subsP. multocida</span> str. HN06, and complete genome. Clustal w multiple sequence alignment was performed for nine sequences showing best hit with sheep isolate <span class="CharOverride-5">Oma87</span> gene sequence and phylogenetic tree was constructed using MEGA 6 (<a href="#Figure-2--P"><span class="Hyperlink">Figure 2</span></a>). O<span class="CharOverride-5">ma87</span> gene from <span class="CharOverride-5">P. multocida</span> (S02)<span class="CharOverride-5"> </span>isolate was found group with <span class="CharOverride-5">P. multocida </span>P52 <span class="CharOverride-5">P. multocid </span>and PMX strains of cattle origin indicating close phylogenetic relationship.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			
          <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117035404.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117035404.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-3-"></a>Figure 3: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117035404.png">3D structure of <span class="CharOverride-5">Oma87</span> protein of <span class="CharOverride-5">P. multocida </span>sheep isolate predicted using Swizz modeller</a></p>
            <p>&nbsp;</p>
          </div>
          <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117032505.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117032505.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-4"></a>Figure 4: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117032505.png">Ramachandran plot for 3D model of the <span class="CharOverride-5">Oma87</span> protein</a></p>
       </div><br>
			
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Table-2-"></a>Table 2: </span>VADAR validation DATA of the predicted 3D model of the <span class="CharOverride-5">Oma87</span> protein</p>
			<table width="657" height="339" class="Table-Style-1 TableOverride-1" id="table-2">
				<colgroup>
					<col class="_idGenTableRowColumn-10" />
					<col class="_idGenTableRowColumn-11" />
					<col class="_idGenTableRowColumn-12" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-22">Statistic</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-22">Observed</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-22">Expected</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Helix</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">107(13%)</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Beta</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">368(47%)</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Coil</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">297(38%)</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Turn</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">96(12%)</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">-</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td colspan="3">
							<p class="Normal" lang="en-US"><span class="Title CharOverride-22">Hydrogen bonds</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Mean HBond distance</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">2.3</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">2.2</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Mean HBond Energy</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">-1.6</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">-2.0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Re with Hbonds</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">525(68%)</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">579(75%)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td colspan="3">
							<p class="Normal" lang="en-US"><span class="Title CharOverride-22">Volume</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Total volume(packing)</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">105946.0</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">104483.6</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-15">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Mean residue volume</span></p>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Mean frac volume</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">137.2</span></p>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">1.1</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">125.0</span></p>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">1.0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">Molecular weight</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">85680.2</span></p>
						</td>
						<td>
							<p class="Normal" lang="en-US"><span class="Title CharOverride-23">-</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Body-Text">&nbsp;</p>
			<p class="Body-Text">&nbsp;</p>
			<p class="Body-Text"><span class="CharOverride-5">Oma87</span> protein sequence was used for protein modelling using SWISS–MODEL homology modelling. From the SWIZZ-MODEL templates library, the template that matched <span class="CharOverride-5">Oma87</span> protein sequence was searched with BLAST and HHBlits and overall 82 templates were found. <span class="CharOverride-5">Oma87</span> protein sequence showed 59.92% identity with outer membrane protein assembly factor BamA structure (protein data bank code 4k3c, chain A) from <span class="CharOverride-5">Haemophilus ducreyi </span>and the same was used as template for model building.3D Model for <span class="CharOverride-5">Oma87</span> protein (<a href="#Figure-3-"><span class="Hyperlink">Figure 3</span></a>) was built based on the target-template alignment using ProMod and then the model quality was assessed using the QMEAN scoring function of -7.64.  The <span class="CharOverride-5">Oma87</span> protein structure was validated using VADAR (<a href="#Table-2-"><span class="Hyperlink">Table 2</span></a>) and RAMACHANDRAN plot (<a href="#Figure-4"><span class="Hyperlink">Figure 4</span></a>). VADAR validation data showed that the predicted protein contained 13% helix, 47% beta, 38% coil, 12% turn and molecular weight of 85.7 kDa. Ramachandran plot statistics obtained from the PROCHECK analysis revealed that 77.4% of amino acids fall in most favoured region, 16% in additionally allowed region, 3.9 % in generously allowed region and 2.7 % in disallowed region. The physiochemical parameters of the <span class="CharOverride-5">Oma87</span> protein were computed using the Expasy’s ProtParam tool. The computed pI value was 5.94 (pI&lt;7) indicates the acidic nature of the <span class="CharOverride-5">Oma87</span> protein. Instability index value is computed to be 30.11 and this classifies the <span class="CharOverride-5">Oma87</span> protein as a sta<span class="Body-Text ParaOverride-1">ble protein. The aliphatic index value is 79.97 shows that <span class="CharOverride-5">Oma87</span> protein may be stable at wide range of temperature. Grand average of hydropathicity index (GRAVY) indicates the solubility of the proteins, for <span class="CharOverride-5">Oma87</span> protein it was found to be -0.387 showing that the protein was hydrophilic in nature. VaxiJen classified <span class="CharOverride-5">Oma87</span> protein as probable antigen because it has a value of 0.5514 which is above the normal threshold value of 0.4, proved that <span class="CharOverride-5">Oma87</span> protein can act as the immunogenic protein<span class="CharOverride-5">.</span></span></p>
			<p class="Body-Text">&nbsp;</p>
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117031006.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117031006.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-5"></a>Figure 5a: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117031006.png">3D structure of MHC class II binding FYQYYSAGGIGSLRG epitope (a</a><br>
			</p>
		  </div>
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117034406.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117034406.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-5"></a>Figure 5b: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117034406.png">MHC class I binding FYQYYSAGGIGSL epitope of <span class="CharOverride-5">Oma87</span> protein (b)</a></p>
       </div><br>
			
		  <p class="Body-Text">&nbsp;</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Table-3-"></a>Table 3</span>: Predicted T cell epitopes of <span class="CharOverride-5">Oma87</span> protein binding with MHC class I alleles and their antigen score<span class="Title CharOverride-25" lang="en-US"><a id="x.30119"></a></span></p>
			<table width="657" height="692" class="Table-Style-1 TableOverride-2" id="table-3">
				<colgroup>
					<col class="_idGenTableRowColumn-17" />
					<col class="_idGenTableRowColumn-18" />
					<col class="_idGenTableRowColumn-19" />
					<col class="_idGenTableRowColumn-20" />
					<col class="_idGenTableRowColumn-21" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">Peptide</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">Position</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">MHC class I Allele</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">IC50</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-7">Vaxijen score</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">VGYTYNKL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">524-531</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.5413</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">DEQDKTISLDFI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">325-336</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kk</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">14</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.8986</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">SYQASIKQDNFL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">433-444</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">16</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.6612</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">GYFPTKGVRANI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">574-585</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">23</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.0916</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">VEAGKSYTVRQI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">337-348</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kk</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">23</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.8904</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">AAYGRTSYGGNL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">497-508</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">27</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.8941</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">RYNAKVEAI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">143-151</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">31</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.03</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">DEIEQFQFSI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">777-786</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kk</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">34</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.5781</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">IGYGTESGL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">424-432</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">36</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.4592</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">SYCLVSSDV</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">672-680</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">43</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.9801</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">RYNAKVEAIVNTL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">143-155</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">44</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.4937</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">VALQWQSPI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">752-760</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">58</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.5112</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">AAYGRTSYGGNLTL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">497-510</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">59</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.9513</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">QERNTGSINFGI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">413-424</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kk</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">70</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.1842</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">KYQGDEIEQFQFSI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">773-786</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">79</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.0124</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">INFGIGYGTESGL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">420-432</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">88</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.1467</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">MELQTDAWWKLF</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">194-205</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kk</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">96</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.9976</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">FDEQDKTISLDFI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">324-336</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kk</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">102</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.0353</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">KEGDLYTVKCARI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">256-268</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kk</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">107</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.7008</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">QYYSAGGIGSL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">640-650</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">121</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.4361</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">GYAQFQILDTDI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">229-240</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">149</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.6808</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">SYQASIKQDNFLGM</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">433-446</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">160</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.4972</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">FYQYYSAGGIGSL</span><span class="Title CharOverride-4">*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">638-650</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">198</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.4882</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">LSYQASIKQDNFL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">432-444</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">200</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.7496</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">QYDDVKASREGNTL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">68-81</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">201</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.3941</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">IPDEAIKQNL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">102-111</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Ld</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">207</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.8153</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">ISASFADGFGGKRL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">623-636</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">210</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.0221</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">IPGSDNKYYKL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">591-601</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Ld</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">221</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.6592</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">AYGRTSYGGNLTL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">498-510</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H-2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">242</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.9579</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-17">*</span><span class="CharOverride-26"> </span><span class="CharOverride-17">Epitope selected for docking studies</span></p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Table-4"></a>Table 4: </span>Predicted T cell epitopes of <span class="CharOverride-5">Oma87</span> protein binding with MHC class II alleles and their antigen score</p>
			<table width="556" height="657" class="Table-Style-1 TableOverride-2" id="table-4">
				<colgroup>
					<col class="_idGenTableRowColumn-22" />
					<col class="_idGenTableRowColumn-23" />
					<col class="_idGenTableRowColumn-24" />
					<col class="_idGenTableRowColumn-12" />
					<col class="_idGenTableRowColumn-25" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-26">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">Peptide</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">Position</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">MHC class II Allele</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">IC-50</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-7">Vaxijen Score</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">FGSTTAFAAPFVVKD</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">11-25</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-Iab</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">40.6</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.5836</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">GSLRGFAYGAIGPNA</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">648-662</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">59.7</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.3174</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">LPFYQYYSAGGIGSL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">636-650</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">75.7</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.5686</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">PFYQYYSAGGIGSLR</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">637-651</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">76.4</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.6307</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">FYQYYSAGGIGSLRG*</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">638-652</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">84.1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.6434</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">LGERGYATAQVNVHP</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">308-322</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">94.8</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.0491</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">GERGYATAQVNVHPT</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">309-323</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">94.9</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.8047</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">STTAFAAPFVVKDIR</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">13-27</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">97.7</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.4277</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">QIRFEGNTSSADSTL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">347-361</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">101</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.4531</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">RQIRFEGNTSSADST</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">346-360</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">105.1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.6825</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">ERGYATAQVNVHPTF</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">310-324</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">105.1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.8262</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">KLGERGYATAQVNVH</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">307-321</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">114.1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.8048</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">VRQIRFEGNTSSADS</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">345-359</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">120.2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.3336</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">VTASTELIVPTPFVA</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">687-701</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">124.9</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.7166</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">TTAFAAPFVVKDIRV</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">14-28</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">127.5</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.7409</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">IRFEGNTSSADSTLR</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">348-362</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">128.4</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.5361</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">RGYATAQVNVHPTFD</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">311-325</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">149.8</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.8189</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">TVRQIRFEGNTSSAD</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">344-358</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">152.2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.2743</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">FEEYDSSKSNTSAAY</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">485-499</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">153.8</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.2213</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">FFEEYDSSKSNTSAA</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">484-498</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">158.6</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.117</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">VFFEEYDSSKSNTSA</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">483-497</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">186.9</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.0261</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">ELIVPTPFVADKNQN</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">692-706</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">188.3</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.4855</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">RLPFYQYYSAGGIGS</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">635-649</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">195.1</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.0154</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">RFEGNTSSADSTLRQ</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">349-363</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">226.3</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.4227</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">NVFFEEYDSSKSNTS</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">482-496</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">238.2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.9596</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">EEYDSSKSNTSAAYG</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">486-500</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">H2-IAb</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">256.3</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-6">1.271</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Normal" lang="en-US"><span class="Title"><a id="x.30286"></a></span></p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-26">* </span><span class="CharOverride-17">Epitope selected for docking studies</span></p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Body-Text">&nbsp;</p>
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117032407.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117032407.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-6"></a>Figure 6a: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117032407.png">3D structure of MHC class I H2-kDa protein (a)</a></p>
		  </div>
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117035107.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117035107.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-6"></a>Figure 6b: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117035107.png">Ramachandran plot (b)</a></p>
       </div><br>
			
			<p class="Body-Text">In order to increase the immune response against the pathogen, activation of Helper T cells and cytotoxic T cells is necessary. Epitope is an antigenic determinant that plays an important role in immunity of an organism (<a href="#Ansari-HR--Raghava-GP--2010-."><span class="Hyperlink">Ansari and Raghava, 2010</span></a>). Epitope that binds with MHC class I and class II alleles are presented to Helper T cells and cytotoxic T cells for their activation. Hence, the prediction of T cell epitopes that binds with both MHC class I and MHC class II alleles will lead to the development of effective vaccine against <span class="CharOverride-5">P. multocida </span>infection. Since prediction of sheep MHC class I and class II allele binding T cell epitope is not possible with the presently availableprediction server, Mouse MHC class I and class II binders were used for this study. Previous study reported that mouse MHC binding T cell epitopes predicted from the Blue tongue virus non-structural protein NS1 showed increase in IFN-<span class="CharOverride-28">γ</span> production and CTL activity in mice, also proved to be consistently immunogenic in sheep infected with blue tongue virus using IFN-<span class="CharOverride-28">γ</span> elispot assay (Rojas et al, 2014).<span class="CharOverride-16"> </span>Prediction of T cell epitope binding with mouse MHC class I alleles (H2KB, H2Dd, H2Kb, H2Kd, H2Kk, H2Ld) and MHC class II alleles (H2IAb, H2IAd, H2IEd) were done by the prediction server. A total of 56 epitopes binding with MHC class I alleles having IC50 value &lt; 250nm was obtained. Likewise, 59 epitopes of MHC class II binders with IC50 value &lt; 250nm was also obtained. Epitopes having binding affinity threshold below 250nm were selected based on the previous study (<a href="#Sakib-MS--Islam-MR--Hasan-AKMM--Nabi-AHMN--2014-."><span class="Hyperlink">Sakib et al., 2014</span></a>). In order to select the best T cell epitope their antigenicity was cross-checked using VaxiJen server, epitope having antigen score above the threshold value of 0.4 were selected, nearly 29 epitopes binding with MHC class I alleles and 26 epitopes binding with MHC class II alleles satisfied both the criteria of IC50 value &lt;250nm and Antigen score &gt;0.4 (<a href="#Table-3-"><span class="Hyperlink">Table 3</span></a> and <a href="#Table-4"><span class="Hyperlink">Table 4</span></a>). Further analysis showed that the MHC class I binding epitope KEGDLYTVKCARI  had the highest antigen score of 1.7 and RQIRFEGNTSSADST epitope had the highest antigen score of 1.6 in the case of MHC class II binders. Best T cell epitope should be able to bind with both MHC class I and MHC class II alleles, out of 29 epitopes from MHC class I H2-kDa binding FYQYYSAGGIGSL and QYYSAGGIGSL epitopes share their sequence with MHC class IIH2-IAb binding FYQYYSAGGIGSLRG and PFYQYYSAGGIGSLR epitopes. From these FYQYYSAGGIGSLRG epitope had higher antigen score of 0.6434 showed sequence similarity with both MHC class I binding FYQYYSAGGIGSL and QYYSAGGIGSL epitopes, whereas FYQYYSAG </p>
			<p class="Body-Text ParaOverride-4">&nbsp;</p>
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117032508.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117032508.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-7"></a>Figure 7a: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117032508.png">3D structure of MHC class II H2-IAb protein (a)</a><br>
       </p>
		  </div>
		  <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117035308.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117035308.png" width="80" height="80"></a>
            
         <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-7"></a>Figure 7b: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117035308.png">Ramachandran plot (b)</a></p>
       </div><br>
			
		  <p class="Body-Text">&nbsp;</p>
		  <p class="Body-Text">GIGSL epitope showed higher antigenic score of 0.4882 compared to the score of QYYSAGGIGSL epitopes (0.432). Based on this FYQYYSAGGIGSLRG and FYQYYSAGGIGSL were selected for further docking studies with MHC class II H2-IAb allele and MHC class I H2-kDa allele. Sequence similarity search of these epitopes was done using BLAST against <span class="CharOverride-5">Mus musculus</span>, <span class="CharOverride-5">Ovis aries</span> and <span class="CharOverride-5">Gallus gallus</span>; no similarity was found which showed the uniqueness of the epitopes. B cell epitopes including linear and conformational structure mostly induce humoral immune response (<a href="#Conway-JF--Watts-NR--Belnap-DM--Cheng-N--Stahl-SJ--Wingfield-PT--Steven-AC--2003-."><span class="Hyperlink">Conway et al., 2003</span></a>; <a href="#Malm-M--Rollman-E--Ustav-M--Hinkula-J--Krohn-K--Wahren-B--Blazevic-V--2005"><span class="Hyperlink">Malm et al., 2005</span></a>). Twenty B cell epitopes of <span class="CharOverride-5">Oma87</span> protein was predicted using ABC prediction server (<a href="#Table-5"><span class="Hyperlink">Table 5</span></a>). Of these EGNTSSADSTLRQEMRQQEG epitope shows high antigenic score of 1.5 and epitope ADGFGGKRLPFYQYYSAGGI with antigen score of 0.8 shares partially similar sequence with FYQYYSAGGIGSLRG and FYQYYSAGGIGSL epitopes.</p>
		  <p class="Body-Text">&nbsp;</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Table-5"></a>Table 5: </span>Predicted B cell epitopes of <span class="CharOverride-5">Oma87</span> protein and their respective antigen score</p>
			<table width="657" height="485" class="Table-Style-1 TableOverride-1" id="table-5">
				<colgroup>
					<col class="_idGenTableRowColumn-27" />
					<col class="_idGenTableRowColumn-28" />
					<col class="_idGenTableRowColumn-29" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-26">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">Position</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-5"><span class="Title CharOverride-4">Epitope</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-4">Score</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-30">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">81</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">LVVTVMPKPVISNVVIDGNK</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.34</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">488</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">YDSSKSNTSAAYGRTSYGGN</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.34</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-30">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">351</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">EGNTSSADSTLRQEMRQQEG</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.59</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">149</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">EAIVNTLPNNSAEIKIQINE</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.897</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-30">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">581</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">VRANIGGRVTIPGSDNKYYK</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.465</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">26</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">IRVDGVQAGTEGSVLATLPV</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.33</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-30">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">384</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">RLDRTGFFESVETKTEAIPG</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.04</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">761</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">GPLVSSYAKPLKKYQGDEIE</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.164</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-30">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">724</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">WKAEDKAKFAKLNVPDYSDP</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.29</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">243</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">SDDKKEARVIIKVKEGDLYT</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.79</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-30">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">313</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">YATAQVNVHPTFDEQDKTIS</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.68</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">412</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">SQERNTGSINFGIGYGTESG</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.28</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-30">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">513</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">PVNENNSYYLGVGYTYNKLK</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.35</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">535</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">APEYNRDLYRQSMKYNDSWT</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.46</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-30">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">602</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">NAEAQGFYPLDREHGWVLSS</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.09</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">680</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">VIGGNAMVTASTELIVPTPF</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.75</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">191</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">EDQMELQTDAWWKLFGNKFD</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.92</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-16">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">628</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">ADGFGGKRLPFYQYYSAGGI</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">0.80</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">445</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">GMGSSISLGGTRNDYGTTIN</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title">1.26</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">659</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-17">GPNAIYRTRQCPDSYCLVSS</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-1"><span class="Title CharOverride-8">0.08</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The 3D structure of the FYQYYSAGGIGSLRG and FYQYYSAGGIGSL epitopes was predicted using PEP fold structure prediction server. Best model proposed by the server was selected for docking studies (<a href="#Figure-5"><span class="Hyperlink">Figure 5a</span></a> and <a href="#Figure-5"><span class="Hyperlink">5b</span></a>). 3D structure of MHC class I H2-KDa allele and MHC class II H2-IAb allele were predicted by Swiss-modeller using the protein sequence obtained from NCBI database AAX98169.1 and NP_996988.2 respectively.3D model of MHC class I H2-KDa (<a href="#Figure-6-"><span class="Hyperlink">Figure 6</span></a>) was built using the template H-2 class I histocompatibility antigen, D-37 alpha chain (3vj6.1). Sequence identity of the template to the MHC class I H2-kDa was 90% and the quality was accessed by Qmean scoring function of -0.83.The <span class="CharOverride-8">template 3K peptide, linker and MHC H-2 class II I-Ab beta chain (3rdt.1.B) was used to build the3D model of MHC class II H2-IAb (</span><span class="Hyperlink">F</span><a href="#Figure-7"><span class="Hyperlink">igure 7</span></a><span class="CharOverride-8">). Sequence identity of the template to the query sequence was 98% and Q mean score was -0.92.3D model of both MHC class II H2-IAb allele and MHC class I H2- kDa allele was validated using VADAR (</span><a href="#Table-6-"><span class="Hyperlink">Table 6</span></a><span class="CharOverride-8">) and RAMACHANDRAN plot (</span><a href="#Figure-6-"><span class="Hyperlink">Figure 6</span></a><span class="CharOverride-8"> and </span><a href="#Figure-7"><span class="Hyperlink">7</span></a><span class="CharOverride-8">).</span></p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Table-6-"></a>Table 6: </span>VADAR validation of 3D structure of MHC class I H2-kDa protein and MHC class II H2-IAb protein</p>
			<table width="657" height="339" class="Table-Style-1 TableOverride-1" id="table-6">
				<colgroup>
					<col class="_idGenTableRowColumn-31" />
					<col class="_idGenTableRowColumn-32" />
					<col class="_idGenTableRowColumn-11" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-33">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">Statistics</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">MHC Class I-H2-Kd</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">MHC Class II-H2-IAb</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Helix</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">0</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">28(14%)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Beta</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">61(61%)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">91(46%)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Coil</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">38(38%)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">76(38%)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Turn</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">24(24%)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">28(14%)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td colspan="3">
							<p class="Body-Text ParaOverride-4"><span class="Title">Hydrogen bonds</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-26">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Mean HBond distance</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title"> 2.2</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">2.2</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Mean HBond Energy</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-1.7</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-1.7</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Re with Hbonds</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title"> 60(60%)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">132(67%)</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td colspan="3">
							<p class="Body-Text ParaOverride-4"><span class="Title">Volume</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-26">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Total volume(packing)</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">13010.3</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">25968.4</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-34">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Mean residue volume</span></p>
							<p class="Body-Text ParaOverride-4"><span class="Title">Mean frac volume</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">131.4</span></p>
							<p class="Body-Text ParaOverride-4"><span class="Title">1.0</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">133.0</span></p>
							<p class="Body-Text ParaOverride-4"><span class="Title">1.0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-13">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Molecular weight</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">11388.26</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">22894.34</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Body-Text">&nbsp;</p>
			<p class="Body-Text">&nbsp;</p>
			<p class="Body-Text">Using PATCHDOCK server binding model of selected T cell epitopes with their respective MHC class I and MHC class II alleles were generated and their refinement was done by FIREDOCK server. FYQYYSAGGIGSLRG epitope was found to bind to the MHC class II H2-IAb allele with the binding score of 5918 and binding energy of -34.09 (<a href="#Figure-8"><span class="Hyperlink">Figure 8</span></a> and <a href="#Table-7"><span class="Hyperlink">Table 7</span></a>). In the case of FYQYYSAGGIGSL epitope-MHC class I H2-kDa allele (<a href="#Figure-9"><span class="Hyperlink">Figure 9</span></a>) the binding score and the binding energy were 8238 and -60.15 respectively (<a href="#Table-7"><span class="Hyperlink">Table 7</span></a>). Based on the binding affinity with MHC class II allele, antigenic score and the docking result FYQYYSAGGIGSLRG epitope which could be superimposed with MHC class I allele binding FYQYYSAGGIGSL epitope may be a suitable vaccine candidate against sheep Pasteurellosis<span class="CharOverride-5">. </span>Longer duration of vaccine- mediated immunity can be achieved by the combination of both humoral and cell mediated immunity (Amanna IJ and Slifka MK, 2011).Hence, we suggest that the B cell epitope ADGFGGKRLPFYQYYSAGGI which had partially similar sequence with predicted T cell epitope can be coupled for inducing antibody response and cell mediated immunity. In this present work, using computational analysis we predicted both T cell and B cell  binding epitopes of <span class="CharOverride-5">Oma87</span> protein with lesser time which may consume more time and  extensive trial if carried out by experimental approaches. Further <span class="CharOverride-5">in vivo</span> work with the predicted epitopes in animals would show their ability to induce protective immune responses against <span class="CharOverride-5">P. multocida.</span></p>
            <p class="Body-Text">&nbsp;</p>
            <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117033609.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117033609.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-8"></a>Figure 8: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117033609.png">Molecular docking of FYQYYSAGGIGSL epitope with MHC class I H2-kDa protein</a></p>
       </div><br>
			
			<div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150117031510.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150117031510.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Figure-9"></a>Figure 9: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150117031510.png">Molecular docking of FYQYYSAGGIGSLRG epitope withMHC class II H2-IAb protein</a></p>
       </div><br>
			
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-4"><a id="Table-7"></a>Table 7: </span>Binding affinity of FYQYYSAGGIGSL epitope with MHC class I H2-kDa protein and FYQYYSAGGIGSLRG epitope with MHC class II H2-IAb protein</p>
			<table width="657" class="Table-Style-1 TableOverride-1" id="table-7">
				<colgroup>
					<col class="_idGenTableRowColumn-1" />
					<col class="_idGenTableRowColumn-35" />
					<col class="_idGenTableRowColumn-36" />
					<col class="_idGenTableRowColumn-37" />
					<col class="_idGenTableRowColumn-38" />
					<col class="_idGenTableRowColumn-39" />
					<col class="_idGenTableRowColumn-40" />
					<col class="_idGenTableRowColumn-41" />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-26">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">Epitope</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">MHC alleles</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">Global energy</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">Binding score</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">Attractive </span></p>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">VdW</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">Repulsive </span></p>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">VdW</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">ACE</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title CharOverride-4">HB</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-14">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">FYQYYSAGGIGSL</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Class I H2-kDa</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-26.72</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">5390</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-26.93</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">12.17</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-5.04</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-2.95</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-26">
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">FYQYYSAGGIGSLRG  </span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">Class II H2-I-AB</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-60.15</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">8238</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-32.03</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">20.38</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-15.50</span></p>
						</td>
						<td>
							<p class="Body-Text ParaOverride-4"><span class="Title">-1.57</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Body-Text">&nbsp;</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Heading-1--Introduction----">ACKNOWLEDGEMENTS</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Sincere thanks are due to ICAR, New Delhi and Dean, Madras Veterinary College, Chennai for providing necessary facilities.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">REFERENCE</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			
			  <li class="References ParaOverride-6" lang="en-US"><a id="Al-Hasani-K--Boyce-J--McCarl-VP--Bottomley-S--Wilkie-I--Adler-B--2007-."></a>Al-Hasani K, Boyce J, McCarl VP, Bottomley S, Wilkie I, Adler B (2007). Identification of novel immunogens in <span class="CharOverride-5">Pasteurella multocida.</span> Microb. Cell Fact. 6(3): 1-12.</li>
				<li class="References ParaOverride-6" lang="en-US"><a id="Adler-R--Adler-B--1991-"></a>Adler R, Adler B (1991). Opsonic monoclonal antibodies against lipopolysaccharide (LPS) antigens of P. multocida and the role of LPS in immunity. Vet. Microbiol. 26(4): 335-347. <a href="http://dx.doi.org/10.1016/0378-1135(91)90027-D"><span class="Hyperlink">http://dx.doi.org/10.1016/0378-1135(91)90027-D</span></a></li>
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