A small derivative of the broad-host-range plasmid RK2 which can be switched from a replicating to a non-replicating state as a response to an externally added inducer
Abstract TrfA is the only plasmid-encoded protein required for RK2 replication. We report here the construction and characterization of an RK2-based vector in which trfA is expressed from the inducible promoter Pm. The resulting construct, pJBSD1, was found to replicate in Escherichia coli DH5a (rec...
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| Veröffentlicht in: | FEMS microbiology letters 1999-11, Vol.180 (2), p.221-227 |
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| creator | Karunakaran, Ponniah Endresen, Dag Terje Ertesvåg, Helga Blatny, Janet Martha Valla, Svein |
| description | Abstract
TrfA is the only plasmid-encoded protein required for RK2 replication. We report here the construction and characterization of an RK2-based vector in which trfA is expressed from the inducible promoter Pm. The resulting construct, pJBSD1, was found to replicate in Escherichia coli DH5a (recA−) only in the presence of a Pm inducer. In two tested E. coli recA+ strains pJBSD1 could replicate in the absence of inducer, but a replication inducer-dependent phenotype was obtained in these strains by introducing a mutation known to reduce the trfA expression level. The plasmid construct could be used as a conditional suicide vector system for targeted chromosomal integration via homologous recombination. This feature may potentially be used for many types of studies in microbial molecular biology. |
| doi_str_mv | 10.1111/j.1574-6968.1999.tb08799.x |
| format | Article |
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TrfA is the only plasmid-encoded protein required for RK2 replication. We report here the construction and characterization of an RK2-based vector in which trfA is expressed from the inducible promoter Pm. The resulting construct, pJBSD1, was found to replicate in Escherichia coli DH5a (recA−) only in the presence of a Pm inducer. In two tested E. coli recA+ strains pJBSD1 could replicate in the absence of inducer, but a replication inducer-dependent phenotype was obtained in these strains by introducing a mutation known to reduce the trfA expression level. The plasmid construct could be used as a conditional suicide vector system for targeted chromosomal integration via homologous recombination. This feature may potentially be used for many types of studies in microbial molecular biology.</description><identifier>ISSN: 0378-1097</identifier><identifier>EISSN: 1574-6968</identifier><identifier>DOI: 10.1111/j.1574-6968.1999.tb08799.x</identifier><identifier>PMID: 10556715</identifier><identifier>CODEN: FMLED7</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Bacteriological methods and techniques used in bacteriology ; Bacteriology ; Biological and medical sciences ; DNA Replication ; E coli ; Escherichia coli - genetics ; Escherichia coli - growth & development ; Escherichia coli Proteins ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation, Bacterial ; Genetic Vectors ; Homologous recombination ; Homology ; Inducer ; Microbiology ; Microorganisms ; Molecular biology ; Mutation ; Phenotypes ; Plasmid ; Plasmids - genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; RecA protein ; Replication ; RK2 ; Suicide ; Suicide vector ; Temperature ; Transformation, Bacterial</subject><ispartof>FEMS microbiology letters, 1999-11, Vol.180 (2), p.221-227</ispartof><rights>1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved. 1999</rights><rights>2000 INIST-CNRS</rights><rights>1999 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-c4571-6af33c353871b8df316fd14fdf92e92beab9f562d50ef4eefcea47393c29910a3</citedby><cites>FETCH-LOGICAL-c4571-6af33c353871b8df316fd14fdf92e92beab9f562d50ef4eefcea47393c29910a3</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.1574-6968.1999.tb08799.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1574-6968.1999.tb08799.x$$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=1185246$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10556715$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Karunakaran, Ponniah</creatorcontrib><creatorcontrib>Endresen, Dag Terje</creatorcontrib><creatorcontrib>Ertesvåg, Helga</creatorcontrib><creatorcontrib>Blatny, Janet Martha</creatorcontrib><creatorcontrib>Valla, Svein</creatorcontrib><title>A small derivative of the broad-host-range plasmid RK2 which can be switched from a replicating to a non-replicating state as a response to an externally added inducer</title><title>FEMS microbiology letters</title><addtitle>FEMS Microbiol Lett</addtitle><description>Abstract
TrfA is the only plasmid-encoded protein required for RK2 replication. We report here the construction and characterization of an RK2-based vector in which trfA is expressed from the inducible promoter Pm. The resulting construct, pJBSD1, was found to replicate in Escherichia coli DH5a (recA−) only in the presence of a Pm inducer. In two tested E. coli recA+ strains pJBSD1 could replicate in the absence of inducer, but a replication inducer-dependent phenotype was obtained in these strains by introducing a mutation known to reduce the trfA expression level. The plasmid construct could be used as a conditional suicide vector system for targeted chromosomal integration via homologous recombination. This feature may potentially be used for many types of studies in microbial molecular biology.</description><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Bacteriological methods and techniques used in bacteriology</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>DNA Replication</subject><subject>E coli</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - growth & development</subject><subject>Escherichia coli Proteins</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Genetic Vectors</subject><subject>Homologous recombination</subject><subject>Homology</subject><subject>Inducer</subject><subject>Microbiology</subject><subject>Microorganisms</subject><subject>Molecular biology</subject><subject>Mutation</subject><subject>Phenotypes</subject><subject>Plasmid</subject><subject>Plasmids - genetics</subject><subject>Polymerase Chain Reaction</subject><subject>Promoter Regions, Genetic</subject><subject>RecA protein</subject><subject>Replication</subject><subject>RK2</subject><subject>Suicide</subject><subject>Suicide vector</subject><subject>Temperature</subject><subject>Transformation, Bacterial</subject><issn>0378-1097</issn><issn>1574-6968</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</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>eNqVkdGK1DAUhoso7rj6ChJUvOuYpE3beCEsi6viiCB6HdLkZCdD29Qk3Zl5Il_TdDvoInphbg4k3_lzDl-WPSN4TdJ5tVsTVpd5xatmTTjn69jipk71cC9b_Xq6n61wUTc5wbw-yx6FsMMYlxRXD7MzghmrasJW2Y8LFHrZdUiDtzcy2htAzqC4BdR6J3W-dSHmXg7XgMZOht5q9OUjRfutVVuk5IBaQGFvo9qCRsa7HknkYeysSmHDNYouXQxuyO9ehigjIBlu2TC6IcAtOCA4RPBDGuiIpNYp0g56UuAfZw-M7AI8OdXz7NvV26-X7_PN53cfLi82uSpZTfJKmqJQBSuamrSNNgWpjCal0YZT4LQF2XLDKqoZBlMCGAWyrAteKMo5wbI4z14uuaN33ycIUfQ2KOg6OYCbgqg4rcuyIQl8_ge4c9M8eRC0IKRmpMIz9XqhlHcheDBi9LaX_igIFrNMsROzMTEbE7NMcZIpDqn56emLqe1B32ld7CXgxQmQQcnOJE3Kht8caRgtq4S9WbC97eD4HxOIq08bSuct2BLgpvEf7fnfFvgJI7TOMA</recordid><startdate>199911</startdate><enddate>199911</enddate><creator>Karunakaran, Ponniah</creator><creator>Endresen, Dag Terje</creator><creator>Ertesvåg, Helga</creator><creator>Blatny, Janet Martha</creator><creator>Valla, Svein</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>199911</creationdate><title>A small derivative of the broad-host-range plasmid RK2 which can be switched from a replicating to a non-replicating state as a response to an externally added inducer</title><author>Karunakaran, Ponniah ; Endresen, Dag Terje ; Ertesvåg, Helga ; Blatny, Janet Martha ; Valla, Svein</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4571-6af33c353871b8df316fd14fdf92e92beab9f562d50ef4eefcea47393c29910a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Bacteriological methods and techniques used in bacteriology</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>DNA Replication</topic><topic>E coli</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - growth & development</topic><topic>Escherichia coli Proteins</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation, Bacterial</topic><topic>Genetic Vectors</topic><topic>Homologous recombination</topic><topic>Homology</topic><topic>Inducer</topic><topic>Microbiology</topic><topic>Microorganisms</topic><topic>Molecular biology</topic><topic>Mutation</topic><topic>Phenotypes</topic><topic>Plasmid</topic><topic>Plasmids - genetics</topic><topic>Polymerase Chain Reaction</topic><topic>Promoter Regions, Genetic</topic><topic>RecA protein</topic><topic>Replication</topic><topic>RK2</topic><topic>Suicide</topic><topic>Suicide vector</topic><topic>Temperature</topic><topic>Transformation, Bacterial</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Karunakaran, Ponniah</creatorcontrib><creatorcontrib>Endresen, Dag Terje</creatorcontrib><creatorcontrib>Ertesvåg, Helga</creatorcontrib><creatorcontrib>Blatny, Janet Martha</creatorcontrib><creatorcontrib>Valla, Svein</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>Karunakaran, Ponniah</au><au>Endresen, Dag Terje</au><au>Ertesvåg, Helga</au><au>Blatny, Janet Martha</au><au>Valla, Svein</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A small derivative of the broad-host-range plasmid RK2 which can be switched from a replicating to a non-replicating state as a response to an externally added inducer</atitle><jtitle>FEMS microbiology letters</jtitle><addtitle>FEMS Microbiol Lett</addtitle><date>1999-11</date><risdate>1999</risdate><volume>180</volume><issue>2</issue><spage>221</spage><epage>227</epage><pages>221-227</pages><issn>0378-1097</issn><eissn>1574-6968</eissn><coden>FMLED7</coden><abstract>Abstract
TrfA is the only plasmid-encoded protein required for RK2 replication. We report here the construction and characterization of an RK2-based vector in which trfA is expressed from the inducible promoter Pm. The resulting construct, pJBSD1, was found to replicate in Escherichia coli DH5a (recA−) only in the presence of a Pm inducer. In two tested E. coli recA+ strains pJBSD1 could replicate in the absence of inducer, but a replication inducer-dependent phenotype was obtained in these strains by introducing a mutation known to reduce the trfA expression level. The plasmid construct could be used as a conditional suicide vector system for targeted chromosomal integration via homologous recombination. This feature may potentially be used for many types of studies in microbial molecular biology.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>10556715</pmid><doi>10.1111/j.1574-6968.1999.tb08799.x</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0378-1097 |
| ispartof | FEMS microbiology letters, 1999-11, Vol.180 (2), p.221-227 |
| issn | 0378-1097 1574-6968 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_69274481 |
| source | MEDLINE; Access via Wiley Online Library; Oxford University Press Journals All Titles (1996-Current); Alma/SFX Local Collection |
| subjects | Bacterial Proteins - genetics Bacterial Proteins - metabolism Bacteriological methods and techniques used in bacteriology Bacteriology Biological and medical sciences DNA Replication E coli Escherichia coli - genetics Escherichia coli - growth & development Escherichia coli Proteins Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Bacterial Genetic Vectors Homologous recombination Homology Inducer Microbiology Microorganisms Molecular biology Mutation Phenotypes Plasmid Plasmids - genetics Polymerase Chain Reaction Promoter Regions, Genetic RecA protein Replication RK2 Suicide Suicide vector Temperature Transformation, Bacterial |
| title | A small derivative of the broad-host-range plasmid RK2 which can be switched from a replicating to a non-replicating state as a response to an externally added inducer |
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