Serotype conversion of a Shigella flexneri candidate vaccine strain via a novel site-specific chromosome-integration system
Abstract Shigella flexneri SFL124 (serotype Y) is a promising live oral vaccine candidate, which has been shown to be safe and immunogenic in human volunteers. To change the serotype of this vaccine strain, we inserted a serotype conversion gene cluster into the chromosome of SFL124 by using a bacte...
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| Veröffentlicht in: | FEMS microbiology letters 1998-09, Vol.166 (1), p.79-87 |
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| creator | Guan, Shukui Verma, Naresh K |
| description | Abstract
Shigella flexneri SFL124 (serotype Y) is a promising live oral vaccine candidate, which has been shown to be safe and immunogenic in human volunteers. To change the serotype of this vaccine strain, we inserted a serotype conversion gene cluster into the chromosome of SFL124 by using a bacteriophage-based site-specific integration system. By cloning an integrase gene (int), an attachment site (attP) and a glucosyl transfer gene cluster from bacteriophage SfX into a suicide vector, and subsequently introducing this construct into S. flexneri SFL124, we obtained a S. flexneri strain (designated SFL1213) expressing the serotype X somatic antigen specificity. The strain retained other characteristics of the parent strain, such as colony shape, growth rate, and Congo red binding property. Stability test showed that the serotype X O-antigen specificity in SFL1213 was 100% stable after being cultured approximately 72 successive hours under non-selective condition. In a mouse pulmonary model, the recombinant strain elicited a significant level of humoral antibodies which recognized the lipopolysaccharide (LPS) of a wild-type S. flexneri serotype X strain. The site-specific insertion system will be useful when stable expression of a cloned single copy gene is desired in the chromosome of S. flexneri vaccine candidate, SFL124. |
| doi_str_mv | 10.1111/j.1574-6968.1998.tb13186.x |
| format | Article |
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Shigella flexneri SFL124 (serotype Y) is a promising live oral vaccine candidate, which has been shown to be safe and immunogenic in human volunteers. To change the serotype of this vaccine strain, we inserted a serotype conversion gene cluster into the chromosome of SFL124 by using a bacteriophage-based site-specific integration system. By cloning an integrase gene (int), an attachment site (attP) and a glucosyl transfer gene cluster from bacteriophage SfX into a suicide vector, and subsequently introducing this construct into S. flexneri SFL124, we obtained a S. flexneri strain (designated SFL1213) expressing the serotype X somatic antigen specificity. The strain retained other characteristics of the parent strain, such as colony shape, growth rate, and Congo red binding property. Stability test showed that the serotype X O-antigen specificity in SFL1213 was 100% stable after being cultured approximately 72 successive hours under non-selective condition. In a mouse pulmonary model, the recombinant strain elicited a significant level of humoral antibodies which recognized the lipopolysaccharide (LPS) of a wild-type S. flexneri serotype X strain. The site-specific insertion system will be useful when stable expression of a cloned single copy gene is desired in the chromosome of S. flexneri vaccine candidate, SFL124.</description><identifier>ISSN: 0378-1097</identifier><identifier>EISSN: 1574-6968</identifier><identifier>DOI: 10.1111/j.1574-6968.1998.tb13186.x</identifier><identifier>PMID: 9741086</identifier><identifier>CODEN: FMLED7</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Animals ; Antibodies ; Antibodies, Bacterial - biosynthesis ; Antigens ; Antigens, Bacterial - genetics ; Attachment Sites, Microbiological - genetics ; attP ; Bacterial Vaccines - genetics ; Bacterial Vaccines - immunology ; Bacteriology ; Bacteriophages - genetics ; Biological and medical sciences ; Chromosomes ; Chromosomes, Bacterial - genetics ; Cloning ; Conversion ; Female ; Fundamental and applied biological sciences. Psychology ; Gene expression ; Genes, Bacterial ; Genetic Techniques ; Genetic Vectors ; Growth rate ; Humans ; Immunization ; Immunogenicity ; int ; Integrase ; Integrases - genetics ; Lipopolysaccharides ; Mice ; Mice, Inbred BALB C ; Microbiology ; Multigene Family ; Phages ; Serotype conversion ; Serotyping ; Shigella flexneri ; Shigella flexneri - classification ; Shigella flexneri - genetics ; Shigella flexneri - immunology ; Stability tests ; Suicide ; Vaccines ; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies ; xis</subject><ispartof>FEMS microbiology letters, 1998-09, Vol.166 (1), p.79-87</ispartof><rights>1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved. 1998</rights><rights>1998 INIST-CNRS</rights><rights>1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4869-a29dfa53116dc11d66356717cc27f65789f4a068d7a72ba605dfd82bd6733cfe3</citedby><cites>FETCH-LOGICAL-c4869-a29dfa53116dc11d66356717cc27f65789f4a068d7a72ba605dfd82bd6733cfe3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1574-6968.1998.tb13186.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1574-6968.1998.tb13186.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2414784$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9741086$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Guan, Shukui</creatorcontrib><creatorcontrib>Verma, Naresh K</creatorcontrib><title>Serotype conversion of a Shigella flexneri candidate vaccine strain via a novel site-specific chromosome-integration system</title><title>FEMS microbiology letters</title><addtitle>FEMS Microbiol Lett</addtitle><description>Abstract
Shigella flexneri SFL124 (serotype Y) is a promising live oral vaccine candidate, which has been shown to be safe and immunogenic in human volunteers. To change the serotype of this vaccine strain, we inserted a serotype conversion gene cluster into the chromosome of SFL124 by using a bacteriophage-based site-specific integration system. By cloning an integrase gene (int), an attachment site (attP) and a glucosyl transfer gene cluster from bacteriophage SfX into a suicide vector, and subsequently introducing this construct into S. flexneri SFL124, we obtained a S. flexneri strain (designated SFL1213) expressing the serotype X somatic antigen specificity. The strain retained other characteristics of the parent strain, such as colony shape, growth rate, and Congo red binding property. Stability test showed that the serotype X O-antigen specificity in SFL1213 was 100% stable after being cultured approximately 72 successive hours under non-selective condition. In a mouse pulmonary model, the recombinant strain elicited a significant level of humoral antibodies which recognized the lipopolysaccharide (LPS) of a wild-type S. flexneri serotype X strain. The site-specific insertion system will be useful when stable expression of a cloned single copy gene is desired in the chromosome of S. flexneri vaccine candidate, SFL124.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Antibodies, Bacterial - biosynthesis</subject><subject>Antigens</subject><subject>Antigens, Bacterial - genetics</subject><subject>Attachment Sites, Microbiological - genetics</subject><subject>attP</subject><subject>Bacterial Vaccines - genetics</subject><subject>Bacterial Vaccines - immunology</subject><subject>Bacteriology</subject><subject>Bacteriophages - genetics</subject><subject>Biological and medical sciences</subject><subject>Chromosomes</subject><subject>Chromosomes, Bacterial - genetics</subject><subject>Cloning</subject><subject>Conversion</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene expression</subject><subject>Genes, Bacterial</subject><subject>Genetic Techniques</subject><subject>Genetic Vectors</subject><subject>Growth rate</subject><subject>Humans</subject><subject>Immunization</subject><subject>Immunogenicity</subject><subject>int</subject><subject>Integrase</subject><subject>Integrases - genetics</subject><subject>Lipopolysaccharides</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Microbiology</subject><subject>Multigene Family</subject><subject>Phages</subject><subject>Serotype conversion</subject><subject>Serotyping</subject><subject>Shigella flexneri</subject><subject>Shigella flexneri - classification</subject><subject>Shigella flexneri - genetics</subject><subject>Shigella flexneri - immunology</subject><subject>Stability tests</subject><subject>Suicide</subject><subject>Vaccines</subject><subject>Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies</subject><subject>xis</subject><issn>0378-1097</issn><issn>1574-6968</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqVkV-L1DAUxYso67j6EYSg4ltr0qT5I-yDLK4KIz6sPodMerOboU3GpB1n8MvbMmUeRBHzch_O7-TewymKFwRXZHpvthVpBCu54rIiSslq2BBKJK8OD4rVWXpYrDAVsiRYicfFk5y3GGNWY35RXCjBCJZ8Vfy8hRSH4w6QjWEPKfsYUHTIoNt7fwddZ5Dr4BAgeWRNaH1rBkB7Y60PgPKQjA9o781kCHEPHcp-gDLvwHrnLbL3KfYxxx5KHwa4S2aYF-RjHqB_WjxypsvwbJmXxbeb91-vP5brLx8-Xb9bl5ZJrkpTq9aZhhLCW0tIyzltuCDC2lo43gipHDOYy1YYUW8Mx03rWllvWi4otQ7oZfH69O8uxe8j5EH3Pts5W4A4Zi2oqkmD2T9BwhummBAT-PI3cBvHFKYQuqaUME4Vbybq7YmyKeacwOld8r1JR02wnovUWz23pee29FykXorUh8n8fFkxbnpoz9aluUl_tegmW9O5ZIL1-YzVjDAh50hXJ-yH7-D4Hwfom89roSZ_c_LHcfcXd_mn838BQK3L9w</recordid><startdate>199809</startdate><enddate>199809</enddate><creator>Guan, Shukui</creator><creator>Verma, Naresh K</creator><general>Blackwell Publishing Ltd</general><general>Blackwell</general><general>Oxford University Press</general><scope>IQODW</scope><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>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>199809</creationdate><title>Serotype conversion of a Shigella flexneri candidate vaccine strain via a novel site-specific chromosome-integration system</title><author>Guan, Shukui ; Verma, Naresh K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4869-a29dfa53116dc11d66356717cc27f65789f4a068d7a72ba605dfd82bd6733cfe3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Antibodies, Bacterial - biosynthesis</topic><topic>Antigens</topic><topic>Antigens, Bacterial - genetics</topic><topic>Attachment Sites, Microbiological - genetics</topic><topic>attP</topic><topic>Bacterial Vaccines - genetics</topic><topic>Bacterial Vaccines - immunology</topic><topic>Bacteriology</topic><topic>Bacteriophages - genetics</topic><topic>Biological and medical sciences</topic><topic>Chromosomes</topic><topic>Chromosomes, Bacterial - genetics</topic><topic>Cloning</topic><topic>Conversion</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene expression</topic><topic>Genes, Bacterial</topic><topic>Genetic Techniques</topic><topic>Genetic Vectors</topic><topic>Growth rate</topic><topic>Humans</topic><topic>Immunization</topic><topic>Immunogenicity</topic><topic>int</topic><topic>Integrase</topic><topic>Integrases - genetics</topic><topic>Lipopolysaccharides</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Microbiology</topic><topic>Multigene Family</topic><topic>Phages</topic><topic>Serotype conversion</topic><topic>Serotyping</topic><topic>Shigella flexneri</topic><topic>Shigella flexneri - classification</topic><topic>Shigella flexneri - genetics</topic><topic>Shigella flexneri - immunology</topic><topic>Stability tests</topic><topic>Suicide</topic><topic>Vaccines</topic><topic>Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies</topic><topic>xis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Guan, Shukui</creatorcontrib><creatorcontrib>Verma, Naresh K</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>FEMS microbiology letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Guan, Shukui</au><au>Verma, Naresh K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Serotype conversion of a Shigella flexneri candidate vaccine strain via a novel site-specific chromosome-integration system</atitle><jtitle>FEMS microbiology letters</jtitle><addtitle>FEMS Microbiol Lett</addtitle><date>1998-09</date><risdate>1998</risdate><volume>166</volume><issue>1</issue><spage>79</spage><epage>87</epage><pages>79-87</pages><issn>0378-1097</issn><eissn>1574-6968</eissn><coden>FMLED7</coden><abstract>Abstract
Shigella flexneri SFL124 (serotype Y) is a promising live oral vaccine candidate, which has been shown to be safe and immunogenic in human volunteers. To change the serotype of this vaccine strain, we inserted a serotype conversion gene cluster into the chromosome of SFL124 by using a bacteriophage-based site-specific integration system. By cloning an integrase gene (int), an attachment site (attP) and a glucosyl transfer gene cluster from bacteriophage SfX into a suicide vector, and subsequently introducing this construct into S. flexneri SFL124, we obtained a S. flexneri strain (designated SFL1213) expressing the serotype X somatic antigen specificity. The strain retained other characteristics of the parent strain, such as colony shape, growth rate, and Congo red binding property. Stability test showed that the serotype X O-antigen specificity in SFL1213 was 100% stable after being cultured approximately 72 successive hours under non-selective condition. In a mouse pulmonary model, the recombinant strain elicited a significant level of humoral antibodies which recognized the lipopolysaccharide (LPS) of a wild-type S. flexneri serotype X strain. The site-specific insertion system will be useful when stable expression of a cloned single copy gene is desired in the chromosome of S. flexneri vaccine candidate, SFL124.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>9741086</pmid><doi>10.1111/j.1574-6968.1998.tb13186.x</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0378-1097 |
| ispartof | FEMS microbiology letters, 1998-09, Vol.166 (1), p.79-87 |
| issn | 0378-1097 1574-6968 |
| language | eng |
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| source | MEDLINE; Access via Wiley Online Library; Oxford University Press Journals All Titles (1996-Current); Alma/SFX Local Collection |
| subjects | Animals Antibodies Antibodies, Bacterial - biosynthesis Antigens Antigens, Bacterial - genetics Attachment Sites, Microbiological - genetics attP Bacterial Vaccines - genetics Bacterial Vaccines - immunology Bacteriology Bacteriophages - genetics Biological and medical sciences Chromosomes Chromosomes, Bacterial - genetics Cloning Conversion Female Fundamental and applied biological sciences. Psychology Gene expression Genes, Bacterial Genetic Techniques Genetic Vectors Growth rate Humans Immunization Immunogenicity int Integrase Integrases - genetics Lipopolysaccharides Mice Mice, Inbred BALB C Microbiology Multigene Family Phages Serotype conversion Serotyping Shigella flexneri Shigella flexneri - classification Shigella flexneri - genetics Shigella flexneri - immunology Stability tests Suicide Vaccines Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies xis |
| title | Serotype conversion of a Shigella flexneri candidate vaccine strain via a novel site-specific chromosome-integration system |
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