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Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish
The outer membrane proteins of the warm water fish pathogen, Aeromonas hydrophila have a role in the virulence of the organism and are potential candidates for vaccine development. In this study, the gene encoding an outer membrane protein designated ompTS was amplified by PCR excluding the region c...
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| Published in: | Protein expression and purification 2007-02, Vol.51 (2), p.303-307 |
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| cited_by | cdi_FETCH-LOGICAL-c448t-8e9557326ca391463205721e096833a5ce4d7ebd3e0ebdf715ffa422691c14313 |
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| cites | cdi_FETCH-LOGICAL-c448t-8e9557326ca391463205721e096833a5ce4d7ebd3e0ebdf715ffa422691c14313 |
| container_end_page | 307 |
| container_issue | 2 |
| container_start_page | 303 |
| container_title | Protein expression and purification |
| container_volume | 51 |
| creator | Khushiramani, Rekha Girisha, S.K. Karunasagar, Indrani Karunasagar, Iddya |
| description | The outer membrane proteins of the warm water fish pathogen,
Aeromonas hydrophila have a role in the virulence of the organism and are potential candidates for vaccine development. In this study, the gene encoding an outer membrane protein designated
ompTS was amplified by PCR excluding the region coding for signal peptide, cloned in pQE 30-UA Vector and expressed using induction with isopropyl thiogalactoside (IPTG). The size of the expressed protein was 37
kDa as estimated by migration in 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE). Polyclonal antibodies were raised in mice and rabbit against the purified protein and the reaction of the antibody was confirmed by Western blotting using the purified protein and
A. hydrophila cultures. The Indian major carp,
Labeo rohita Hamilton was immunized using the purified protein and developed antibodies with mean titers of 1:4000 on day 14 and 1:12,000 on day 28 showing promise that the protein is highly immunogenic in fish. |
| doi_str_mv | 10.1016/j.pep.2006.07.021 |
| format | article |
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Aeromonas hydrophila have a role in the virulence of the organism and are potential candidates for vaccine development. In this study, the gene encoding an outer membrane protein designated
ompTS was amplified by PCR excluding the region coding for signal peptide, cloned in pQE 30-UA Vector and expressed using induction with isopropyl thiogalactoside (IPTG). The size of the expressed protein was 37
kDa as estimated by migration in 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE). Polyclonal antibodies were raised in mice and rabbit against the purified protein and the reaction of the antibody was confirmed by Western blotting using the purified protein and
A. hydrophila cultures. The Indian major carp,
Labeo rohita Hamilton was immunized using the purified protein and developed antibodies with mean titers of 1:4000 on day 14 and 1:12,000 on day 28 showing promise that the protein is highly immunogenic in fish.</description><identifier>ISSN: 1046-5928</identifier><identifier>EISSN: 1096-0279</identifier><identifier>DOI: 10.1016/j.pep.2006.07.021</identifier><identifier>PMID: 16959494</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Aeromonas hydrophila ; Aeromonas hydrophila - genetics ; Aeromonas hydrophila - pathogenicity ; Animals ; Bacterial Outer Membrane Proteins - biosynthesis ; Bacterial Outer Membrane Proteins - genetics ; Bacterial Outer Membrane Proteins - immunology ; Carps ; Cloning expression ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Fish Diseases - immunology ; Fish Diseases - prevention & control ; Immunogenicity ; Labeo rohita ; Mice ; Outer membrane protein ; Polyclonal antibodies ; Rabbits</subject><ispartof>Protein expression and purification, 2007-02, Vol.51 (2), p.303-307</ispartof><rights>2006 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c448t-8e9557326ca391463205721e096833a5ce4d7ebd3e0ebdf715ffa422691c14313</citedby><cites>FETCH-LOGICAL-c448t-8e9557326ca391463205721e096833a5ce4d7ebd3e0ebdf715ffa422691c14313</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16959494$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Khushiramani, Rekha</creatorcontrib><creatorcontrib>Girisha, S.K.</creatorcontrib><creatorcontrib>Karunasagar, Indrani</creatorcontrib><creatorcontrib>Karunasagar, Iddya</creatorcontrib><title>Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish</title><title>Protein expression and purification</title><addtitle>Protein Expr Purif</addtitle><description>The outer membrane proteins of the warm water fish pathogen,
Aeromonas hydrophila have a role in the virulence of the organism and are potential candidates for vaccine development. In this study, the gene encoding an outer membrane protein designated
ompTS was amplified by PCR excluding the region coding for signal peptide, cloned in pQE 30-UA Vector and expressed using induction with isopropyl thiogalactoside (IPTG). The size of the expressed protein was 37
kDa as estimated by migration in 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE). Polyclonal antibodies were raised in mice and rabbit against the purified protein and the reaction of the antibody was confirmed by Western blotting using the purified protein and
A. hydrophila cultures. The Indian major carp,
Labeo rohita Hamilton was immunized using the purified protein and developed antibodies with mean titers of 1:4000 on day 14 and 1:12,000 on day 28 showing promise that the protein is highly immunogenic in fish.</description><subject>Aeromonas hydrophila</subject><subject>Aeromonas hydrophila - genetics</subject><subject>Aeromonas hydrophila - pathogenicity</subject><subject>Animals</subject><subject>Bacterial Outer Membrane Proteins - biosynthesis</subject><subject>Bacterial Outer Membrane Proteins - genetics</subject><subject>Bacterial Outer Membrane Proteins - immunology</subject><subject>Carps</subject><subject>Cloning expression</subject><subject>Cloning, Molecular</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Fish Diseases - immunology</subject><subject>Fish Diseases - prevention & control</subject><subject>Immunogenicity</subject><subject>Labeo rohita</subject><subject>Mice</subject><subject>Outer membrane protein</subject><subject>Polyclonal antibodies</subject><subject>Rabbits</subject><issn>1046-5928</issn><issn>1096-0279</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><recordid>eNqFkU9v1DAQxS0EoqXwAbignLgleGzHicWpWvGnUiUOlLPldSZdr2I72AnqfnscdiVucPFYo997mplHyFugDVCQH47NjHPDKJUN7RrK4Bm5BqpkTVmnnm9_IetWsf6KvMr5SCmApO1LcgVStUoocU3W3RSDC4-VCUOFT3PCnF0MVRxLp4rrgqny6PfJBKzmFBd0pe3nh-8bcosp-hhMrg6nIcX54Cbzxykv63DaCOf9GuIjBmfdcqqKeHT58Jq8GM2U8c2l3pAfnz897L7W99--3O1u72srRL_UPaq27TiT1nAFQnJG244BlhV7zk1rUQwd7geOtLxjB-04GsGYVGBBcOA35P3Zt0z-c8W8aO-yxWkq28Q162IjuVL9f0FQknZciQLCGbQp5pxw1HNy3qSTBqq3UPRRl1D0FoqmnS6hFM27i_m69zj8VVxSKMDHM4DlFr8cJp2tw2BxcAntoofo_mH_G0icndg</recordid><startdate>20070201</startdate><enddate>20070201</enddate><creator>Khushiramani, Rekha</creator><creator>Girisha, S.K.</creator><creator>Karunasagar, Indrani</creator><creator>Karunasagar, Iddya</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>20070201</creationdate><title>Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish</title><author>Khushiramani, Rekha ; Girisha, S.K. ; Karunasagar, Indrani ; Karunasagar, Iddya</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c448t-8e9557326ca391463205721e096833a5ce4d7ebd3e0ebdf715ffa422691c14313</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Aeromonas hydrophila</topic><topic>Aeromonas hydrophila - genetics</topic><topic>Aeromonas hydrophila - pathogenicity</topic><topic>Animals</topic><topic>Bacterial Outer Membrane Proteins - biosynthesis</topic><topic>Bacterial Outer Membrane Proteins - genetics</topic><topic>Bacterial Outer Membrane Proteins - immunology</topic><topic>Carps</topic><topic>Cloning expression</topic><topic>Cloning, Molecular</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Fish Diseases - immunology</topic><topic>Fish Diseases - prevention & control</topic><topic>Immunogenicity</topic><topic>Labeo rohita</topic><topic>Mice</topic><topic>Outer membrane protein</topic><topic>Polyclonal antibodies</topic><topic>Rabbits</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Khushiramani, Rekha</creatorcontrib><creatorcontrib>Girisha, S.K.</creatorcontrib><creatorcontrib>Karunasagar, Indrani</creatorcontrib><creatorcontrib>Karunasagar, Iddya</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Protein expression and purification</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Khushiramani, Rekha</au><au>Girisha, S.K.</au><au>Karunasagar, Indrani</au><au>Karunasagar, Iddya</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish</atitle><jtitle>Protein expression and purification</jtitle><addtitle>Protein Expr Purif</addtitle><date>2007-02-01</date><risdate>2007</risdate><volume>51</volume><issue>2</issue><spage>303</spage><epage>307</epage><pages>303-307</pages><issn>1046-5928</issn><eissn>1096-0279</eissn><abstract>The outer membrane proteins of the warm water fish pathogen,
Aeromonas hydrophila have a role in the virulence of the organism and are potential candidates for vaccine development. In this study, the gene encoding an outer membrane protein designated
ompTS was amplified by PCR excluding the region coding for signal peptide, cloned in pQE 30-UA Vector and expressed using induction with isopropyl thiogalactoside (IPTG). The size of the expressed protein was 37
kDa as estimated by migration in 10% sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE). Polyclonal antibodies were raised in mice and rabbit against the purified protein and the reaction of the antibody was confirmed by Western blotting using the purified protein and
A. hydrophila cultures. The Indian major carp,
Labeo rohita Hamilton was immunized using the purified protein and developed antibodies with mean titers of 1:4000 on day 14 and 1:12,000 on day 28 showing promise that the protein is highly immunogenic in fish.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>16959494</pmid><doi>10.1016/j.pep.2006.07.021</doi><tpages>5</tpages></addata></record> |
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| ispartof | Protein expression and purification, 2007-02, Vol.51 (2), p.303-307 |
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| subjects | Aeromonas hydrophila Aeromonas hydrophila - genetics Aeromonas hydrophila - pathogenicity Animals Bacterial Outer Membrane Proteins - biosynthesis Bacterial Outer Membrane Proteins - genetics Bacterial Outer Membrane Proteins - immunology Carps Cloning expression Cloning, Molecular Enzyme-Linked Immunosorbent Assay Fish Diseases - immunology Fish Diseases - prevention & control Immunogenicity Labeo rohita Mice Outer membrane protein Polyclonal antibodies Rabbits |
| title | Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish |
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