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Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species
We recently identified and described a putative prophage on the genomic island FhaGI-1 located within the genome of AS02-814 ( subsp. -like 3523). In this study, we constructed two variants of a phage integration vector, called pFIV1-Val and pFIV2-Val ( Integration Vector-tRNA -specific), using the...
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| Published in: | Frontiers in cellular and infection microbiology 2018-03, Vol.8, p.75-75 |
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| container_title | Frontiers in cellular and infection microbiology |
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| creator | Tlapák, Hana Köppen, Kristin Rydzewski, Kerstin Grunow, Roland Heuner, Klaus |
| description | We recently identified and described a putative prophage on the genomic island FhaGI-1 located within the genome of
AS02-814 (
subsp.
-like 3523). In this study, we constructed two variants of a
phage integration vector, called pFIV1-Val and pFIV2-Val (
Integration Vector-tRNA
-specific), using the
sites and the site-specific integrase (FN3523_1033) of FhaGI-1, a chloramphenicol resistance cassette and a
gene for counter selection of transformants against the vector backbone. We inserted the respective sites and genes into vector pUC57-Kana to allow for propagation in
. The constructs generated a circular episomal form in
which could be used to transform
. where FIV-Val stably integrated site specifically into the tRNA
gene of the genome, whereas pUC57-Kana is lost due to counter selection. Functionality of the new vector was demonstrated by the successfully complementation of a
mutant strain. The vectors were stable
and during host-cell infection without selective pressure. Thus, the vectors can be applied as a further genetic tool in
research, expanding the present genetic tools by an integrative element. This new element is suitable to perform long-term experiments with different
species. |
| doi_str_mv | 10.3389/fcimb.2018.00075 |
| format | article |
| fullrecord | <record><control><sourceid>proquest_doaj_</sourceid><recordid>TN_cdi_doaj_primary_oai_doaj_org_article_3c1117ae7b334dd4b41693cd8a3a0aba</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><doaj_id>oai_doaj_org_article_3c1117ae7b334dd4b41693cd8a3a0aba</doaj_id><sourcerecordid>2019806324</sourcerecordid><originalsourceid>FETCH-LOGICAL-c462t-c69605ec5726f7a801a82c073399db120d45a6dd529d5569c3485712e91533703</originalsourceid><addsrcrecordid>eNpVkc1vEzEQxS1ERavSOyfkI5cN_lh77QsSCg1EqqBS21zNrD2butqsg70B8d-zSUrVnvwx7_0840fIO85mUhr7sfNx084E42bGGGvUK3ImhFSVsMa8frY_JRelPLC9hglj5RtyKqyyNdPmjPycp6GMeefHmAaaOgr0O_6h1_ewRrocRlxnOJRW6MeU6XaxXFUr6Gk3He4K0jjQL7HrMOMw0kWGwceCfQ_0Zos-YnlLTjroC148rufkbnF5O_9WXf34upx_vqp8rcVYeW01U-hVI3TXgGEcjPCskdLa0HLBQq1Ah6CEDUpp62VtVMMFWq6kbJg8J8sjNyR4cNscN5D_ugTRHS5SXjvIY_Q9Ouk55w1g00pZh1C3NddW-mBAAoMWJtanI2u7azcY_DRahv4F9GVliPdunX47ZTTn0kyAD4-AnH7tsIxuE4vff8uAaVfcFJo1TEtRT1J2lPqcSsnYPT3Dmdvn7A457y3GHXKeLO-ft_dk-J-q_AcE9qOB</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2019806324</pqid></control><display><type>article</type><title>Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species</title><source>PubMed Central</source><creator>Tlapák, Hana ; Köppen, Kristin ; Rydzewski, Kerstin ; Grunow, Roland ; Heuner, Klaus</creator><creatorcontrib>Tlapák, Hana ; Köppen, Kristin ; Rydzewski, Kerstin ; Grunow, Roland ; Heuner, Klaus</creatorcontrib><description>We recently identified and described a putative prophage on the genomic island FhaGI-1 located within the genome of
AS02-814 (
subsp.
-like 3523). In this study, we constructed two variants of a
phage integration vector, called pFIV1-Val and pFIV2-Val (
Integration Vector-tRNA
-specific), using the
sites and the site-specific integrase (FN3523_1033) of FhaGI-1, a chloramphenicol resistance cassette and a
gene for counter selection of transformants against the vector backbone. We inserted the respective sites and genes into vector pUC57-Kana to allow for propagation in
. The constructs generated a circular episomal form in
which could be used to transform
. where FIV-Val stably integrated site specifically into the tRNA
gene of the genome, whereas pUC57-Kana is lost due to counter selection. Functionality of the new vector was demonstrated by the successfully complementation of a
mutant strain. The vectors were stable
and during host-cell infection without selective pressure. Thus, the vectors can be applied as a further genetic tool in
research, expanding the present genetic tools by an integrative element. This new element is suitable to perform long-term experiments with different
species.</description><identifier>ISSN: 2235-2988</identifier><identifier>EISSN: 2235-2988</identifier><identifier>DOI: 10.3389/fcimb.2018.00075</identifier><identifier>PMID: 29594068</identifier><language>eng</language><publisher>Switzerland: Frontiers Media S.A</publisher><subject>Bacteriophages - genetics ; Chloramphenicol Resistance - genetics ; DNA, Bacterial - genetics ; Drug Resistance, Bacterial - genetics ; episomal ; Escherichia coli - genetics ; Francisella - genetics ; Francisella - growth & development ; Francisella - virology ; Francisella tularensis ; Francisella tularensis - genetics ; Genetic Vectors ; genomic island ; Genomic Islands ; Humans ; Integrases - genetics ; integrative vector ; Microbiology ; Mutation ; pFIV-Val ; phage ; Plasmids ; Recombination, Genetic ; RNA, Transfer, Val - genetics ; Transformation, Bacterial ; U937 Cells</subject><ispartof>Frontiers in cellular and infection microbiology, 2018-03, Vol.8, p.75-75</ispartof><rights>Copyright © 2018 Tlapák, Köppen, Rydzewski, Grunow and Heuner. 2018 Tlapák, Köppen, Rydzewski, Grunow and Heuner</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c462t-c69605ec5726f7a801a82c073399db120d45a6dd529d5569c3485712e91533703</citedby><cites>FETCH-LOGICAL-c462t-c69605ec5726f7a801a82c073399db120d45a6dd529d5569c3485712e91533703</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5861138/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5861138/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29594068$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tlapák, Hana</creatorcontrib><creatorcontrib>Köppen, Kristin</creatorcontrib><creatorcontrib>Rydzewski, Kerstin</creatorcontrib><creatorcontrib>Grunow, Roland</creatorcontrib><creatorcontrib>Heuner, Klaus</creatorcontrib><title>Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species</title><title>Frontiers in cellular and infection microbiology</title><addtitle>Front Cell Infect Microbiol</addtitle><description>We recently identified and described a putative prophage on the genomic island FhaGI-1 located within the genome of
AS02-814 (
subsp.
-like 3523). In this study, we constructed two variants of a
phage integration vector, called pFIV1-Val and pFIV2-Val (
Integration Vector-tRNA
-specific), using the
sites and the site-specific integrase (FN3523_1033) of FhaGI-1, a chloramphenicol resistance cassette and a
gene for counter selection of transformants against the vector backbone. We inserted the respective sites and genes into vector pUC57-Kana to allow for propagation in
. The constructs generated a circular episomal form in
which could be used to transform
. where FIV-Val stably integrated site specifically into the tRNA
gene of the genome, whereas pUC57-Kana is lost due to counter selection. Functionality of the new vector was demonstrated by the successfully complementation of a
mutant strain. The vectors were stable
and during host-cell infection without selective pressure. Thus, the vectors can be applied as a further genetic tool in
research, expanding the present genetic tools by an integrative element. This new element is suitable to perform long-term experiments with different
species.</description><subject>Bacteriophages - genetics</subject><subject>Chloramphenicol Resistance - genetics</subject><subject>DNA, Bacterial - genetics</subject><subject>Drug Resistance, Bacterial - genetics</subject><subject>episomal</subject><subject>Escherichia coli - genetics</subject><subject>Francisella - genetics</subject><subject>Francisella - growth & development</subject><subject>Francisella - virology</subject><subject>Francisella tularensis</subject><subject>Francisella tularensis - genetics</subject><subject>Genetic Vectors</subject><subject>genomic island</subject><subject>Genomic Islands</subject><subject>Humans</subject><subject>Integrases - genetics</subject><subject>integrative vector</subject><subject>Microbiology</subject><subject>Mutation</subject><subject>pFIV-Val</subject><subject>phage</subject><subject>Plasmids</subject><subject>Recombination, Genetic</subject><subject>RNA, Transfer, Val - genetics</subject><subject>Transformation, Bacterial</subject><subject>U937 Cells</subject><issn>2235-2988</issn><issn>2235-2988</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNpVkc1vEzEQxS1ERavSOyfkI5cN_lh77QsSCg1EqqBS21zNrD2butqsg70B8d-zSUrVnvwx7_0840fIO85mUhr7sfNx084E42bGGGvUK3ImhFSVsMa8frY_JRelPLC9hglj5RtyKqyyNdPmjPycp6GMeefHmAaaOgr0O_6h1_ewRrocRlxnOJRW6MeU6XaxXFUr6Gk3He4K0jjQL7HrMOMw0kWGwceCfQ_0Zos-YnlLTjroC148rufkbnF5O_9WXf34upx_vqp8rcVYeW01U-hVI3TXgGEcjPCskdLa0HLBQq1Ah6CEDUpp62VtVMMFWq6kbJg8J8sjNyR4cNscN5D_ugTRHS5SXjvIY_Q9Ouk55w1g00pZh1C3NddW-mBAAoMWJtanI2u7azcY_DRahv4F9GVliPdunX47ZTTn0kyAD4-AnH7tsIxuE4vff8uAaVfcFJo1TEtRT1J2lPqcSsnYPT3Dmdvn7A457y3GHXKeLO-ft_dk-J-q_AcE9qOB</recordid><startdate>20180314</startdate><enddate>20180314</enddate><creator>Tlapák, Hana</creator><creator>Köppen, Kristin</creator><creator>Rydzewski, Kerstin</creator><creator>Grunow, Roland</creator><creator>Heuner, Klaus</creator><general>Frontiers Media S.A</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>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20180314</creationdate><title>Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species</title><author>Tlapák, Hana ; Köppen, Kristin ; Rydzewski, Kerstin ; Grunow, Roland ; Heuner, Klaus</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c462t-c69605ec5726f7a801a82c073399db120d45a6dd529d5569c3485712e91533703</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Bacteriophages - genetics</topic><topic>Chloramphenicol Resistance - genetics</topic><topic>DNA, Bacterial - genetics</topic><topic>Drug Resistance, Bacterial - genetics</topic><topic>episomal</topic><topic>Escherichia coli - genetics</topic><topic>Francisella - genetics</topic><topic>Francisella - growth & development</topic><topic>Francisella - virology</topic><topic>Francisella tularensis</topic><topic>Francisella tularensis - genetics</topic><topic>Genetic Vectors</topic><topic>genomic island</topic><topic>Genomic Islands</topic><topic>Humans</topic><topic>Integrases - genetics</topic><topic>integrative vector</topic><topic>Microbiology</topic><topic>Mutation</topic><topic>pFIV-Val</topic><topic>phage</topic><topic>Plasmids</topic><topic>Recombination, Genetic</topic><topic>RNA, Transfer, Val - genetics</topic><topic>Transformation, Bacterial</topic><topic>U937 Cells</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tlapák, Hana</creatorcontrib><creatorcontrib>Köppen, Kristin</creatorcontrib><creatorcontrib>Rydzewski, Kerstin</creatorcontrib><creatorcontrib>Grunow, Roland</creatorcontrib><creatorcontrib>Heuner, Klaus</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Frontiers in cellular and infection microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tlapák, Hana</au><au>Köppen, Kristin</au><au>Rydzewski, Kerstin</au><au>Grunow, Roland</au><au>Heuner, Klaus</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species</atitle><jtitle>Frontiers in cellular and infection microbiology</jtitle><addtitle>Front Cell Infect Microbiol</addtitle><date>2018-03-14</date><risdate>2018</risdate><volume>8</volume><spage>75</spage><epage>75</epage><pages>75-75</pages><issn>2235-2988</issn><eissn>2235-2988</eissn><abstract>We recently identified and described a putative prophage on the genomic island FhaGI-1 located within the genome of
AS02-814 (
subsp.
-like 3523). In this study, we constructed two variants of a
phage integration vector, called pFIV1-Val and pFIV2-Val (
Integration Vector-tRNA
-specific), using the
sites and the site-specific integrase (FN3523_1033) of FhaGI-1, a chloramphenicol resistance cassette and a
gene for counter selection of transformants against the vector backbone. We inserted the respective sites and genes into vector pUC57-Kana to allow for propagation in
. The constructs generated a circular episomal form in
which could be used to transform
. where FIV-Val stably integrated site specifically into the tRNA
gene of the genome, whereas pUC57-Kana is lost due to counter selection. Functionality of the new vector was demonstrated by the successfully complementation of a
mutant strain. The vectors were stable
and during host-cell infection without selective pressure. Thus, the vectors can be applied as a further genetic tool in
research, expanding the present genetic tools by an integrative element. This new element is suitable to perform long-term experiments with different
species.</abstract><cop>Switzerland</cop><pub>Frontiers Media S.A</pub><pmid>29594068</pmid><doi>10.3389/fcimb.2018.00075</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Frontiers in cellular and infection microbiology, 2018-03, Vol.8, p.75-75 |
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| subjects | Bacteriophages - genetics Chloramphenicol Resistance - genetics DNA, Bacterial - genetics Drug Resistance, Bacterial - genetics episomal Escherichia coli - genetics Francisella - genetics Francisella - growth & development Francisella - virology Francisella tularensis Francisella tularensis - genetics Genetic Vectors genomic island Genomic Islands Humans Integrases - genetics integrative vector Microbiology Mutation pFIV-Val phage Plasmids Recombination, Genetic RNA, Transfer, Val - genetics Transformation, Bacterial U937 Cells |
| title | Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species |
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