Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants
A negative selectable marker gene, codA, was successfully co-transformed with a GUS reporter gene to develop selectable marker gene-free transgenic plants. The pNC binary vector contained a T-DNA harboring the codA gene next to the nptII gene, while a second binary vector, pHG, contained a GUS repor...
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| Veröffentlicht in: | Theoretical and applied genetics 2004-11, Vol.109 (8), p.1562-1567 |
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| creator | Park, J Lee, Y.K Kang, B.K Chung, W.I |
| description | A negative selectable marker gene, codA, was successfully co-transformed with a GUS reporter gene to develop selectable marker gene-free transgenic plants. The pNC binary vector contained a T-DNA harboring the codA gene next to the nptII gene, while a second binary vector, pHG, contained a GUS reporter gene. Tobacco plants (Nicotiana tabacum cv. Samsun NN) were co-transformed via the mixture method with Agrobacterium tumefaciens LBA4404 strains harboring pNC and pHG, respectively. Seeds harvested from the co-transformants were sown on germination media containing 5-fluorocytosine (5-FC). Analysis of the progeny by GUS staining and PCR amplification revealed that all of the 5-FC-resistant R1 plants were codA free, and that the codA gene segregated independently of the GUS gene. Because codA-free seedlings developed normally on 5-FC-containing medium, we suggest that co-transformation with negatively selectable markers is a viable method for the production of easily distinguished, selectable marker gene-free transgenic plants. |
| doi_str_mv | 10.1007/s00122-004-1790-x |
| format | Article |
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The pNC binary vector contained a T-DNA harboring the codA gene next to the nptII gene, while a second binary vector, pHG, contained a GUS reporter gene. Tobacco plants (Nicotiana tabacum cv. Samsun NN) were co-transformed via the mixture method with Agrobacterium tumefaciens LBA4404 strains harboring pNC and pHG, respectively. Seeds harvested from the co-transformants were sown on germination media containing 5-fluorocytosine (5-FC). Analysis of the progeny by GUS staining and PCR amplification revealed that all of the 5-FC-resistant R1 plants were codA free, and that the codA gene segregated independently of the GUS gene. Because codA-free seedlings developed normally on 5-FC-containing medium, we suggest that co-transformation with negatively selectable markers is a viable method for the production of easily distinguished, selectable marker gene-free transgenic plants.</description><identifier>ISSN: 0040-5752</identifier><identifier>EISSN: 1432-2242</identifier><identifier>DOI: 10.1007/s00122-004-1790-x</identifier><identifier>PMID: 15448898</identifier><identifier>CODEN: THAGA6</identifier><language>eng</language><publisher>Heidelberg: Springer</publisher><subject>5-fluorocytosine ; Agrobacterium tumefaciens ; beta-glucuronidase ; Biological and medical sciences ; Blotting, Southern ; Classical genetics, quantitative genetics, hybrids ; codA gene ; cytosine deaminase ; DNA Primers ; DNA, Bacterial - genetics ; drug resistance ; flucytosine ; Fundamental and applied biological sciences. Psychology ; Genes ; Genetic Engineering - methods ; genetic markers ; Genetic Markers - genetics ; genetic transformation ; Genetic Vectors ; Genetics of eukaryotes. Biological and molecular evolution ; in vitro selection ; negative selection ; Nicotiana - genetics ; Nicotiana tabacum ; nptII gene ; Plants, Genetically Modified ; plasmid vectors ; Polymerase Chain Reaction ; Pteridophyta, spermatophyta ; reporter genes ; seed germination ; tobacco ; Transformation, Genetic - genetics ; Transgenic plants ; Vegetals</subject><ispartof>Theoretical and applied genetics, 2004-11, Vol.109 (8), p.1562-1567</ispartof><rights>2005 INIST-CNRS</rights><rights>Springer-Verlag 2004</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c411t-ed8edd9beb5ec3dbfd36ac7ccdd43e54dec3945e67c2abb99a15899b3a001ec63</citedby><cites>FETCH-LOGICAL-c411t-ed8edd9beb5ec3dbfd36ac7ccdd43e54dec3945e67c2abb99a15899b3a001ec63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=16292977$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15448898$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Park, J</creatorcontrib><creatorcontrib>Lee, Y.K</creatorcontrib><creatorcontrib>Kang, B.K</creatorcontrib><creatorcontrib>Chung, W.I</creatorcontrib><title>Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants</title><title>Theoretical and applied genetics</title><addtitle>Theor Appl Genet</addtitle><description>A negative selectable marker gene, codA, was successfully co-transformed with a GUS reporter gene to develop selectable marker gene-free transgenic plants. The pNC binary vector contained a T-DNA harboring the codA gene next to the nptII gene, while a second binary vector, pHG, contained a GUS reporter gene. Tobacco plants (Nicotiana tabacum cv. Samsun NN) were co-transformed via the mixture method with Agrobacterium tumefaciens LBA4404 strains harboring pNC and pHG, respectively. Seeds harvested from the co-transformants were sown on germination media containing 5-fluorocytosine (5-FC). Analysis of the progeny by GUS staining and PCR amplification revealed that all of the 5-FC-resistant R1 plants were codA free, and that the codA gene segregated independently of the GUS gene. Because codA-free seedlings developed normally on 5-FC-containing medium, we suggest that co-transformation with negatively selectable markers is a viable method for the production of easily distinguished, selectable marker gene-free transgenic plants.</description><subject>5-fluorocytosine</subject><subject>Agrobacterium tumefaciens</subject><subject>beta-glucuronidase</subject><subject>Biological and medical sciences</subject><subject>Blotting, Southern</subject><subject>Classical genetics, quantitative genetics, hybrids</subject><subject>codA gene</subject><subject>cytosine deaminase</subject><subject>DNA Primers</subject><subject>DNA, Bacterial - genetics</subject><subject>drug resistance</subject><subject>flucytosine</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes</subject><subject>Genetic Engineering - methods</subject><subject>genetic markers</subject><subject>Genetic Markers - genetics</subject><subject>genetic transformation</subject><subject>Genetic Vectors</subject><subject>Genetics of eukaryotes. Biological and molecular evolution</subject><subject>in vitro selection</subject><subject>negative selection</subject><subject>Nicotiana - genetics</subject><subject>Nicotiana tabacum</subject><subject>nptII gene</subject><subject>Plants, Genetically Modified</subject><subject>plasmid vectors</subject><subject>Polymerase Chain Reaction</subject><subject>Pteridophyta, spermatophyta</subject><subject>reporter genes</subject><subject>seed germination</subject><subject>tobacco</subject><subject>Transformation, Genetic - genetics</subject><subject>Transgenic plants</subject><subject>Vegetals</subject><issn>0040-5752</issn><issn>1432-2242</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqFkU9v1DAQxS0EotvCB-ACFhLcXMb_4viIVlCQKnEoPVuOPVlSsvFiJ6gc-t3xdleqhFRxsjz-vZnxe4S84nDOAcyHAsCFYACKcWOB3T4hK66kYEIo8ZSs6gMwbbQ4Iael3ACA0CCfkxOulWpb267I3TqxOfup9Clv_TykiS5lmDbU0wk3tfAbacERw-y7EenW55-Y6QYnpFVB5x9IdznFJdxLU_8IzPqMSO8H1esQ6G7001xekGe9Hwu-PJ5n5Przp-_rL-zy28XX9cdLFhTnM8PYYoy2w05jkLHro2x8MCHEqCRqFWvVKo2NCcJ3nbWe69baTvpqEIZGnpH3h751118LltlthxJwrEtgWoprDLSqWvNfkBtlpYZ9x7f_gDdpyVP9hGsFWMlbYSrED1DIqZSMvdvlobryx3Fw-wTdIUFXg3L7BN1t1bw-Nl66LcYHxTGyCrw7Ar4EP_bV0zCUB64RVlizH_7mwPU-Ob_Jlbm-EsAlgG20ViD_ArI0sAw</recordid><startdate>20041101</startdate><enddate>20041101</enddate><creator>Park, J</creator><creator>Lee, Y.K</creator><creator>Kang, B.K</creator><creator>Chung, W.I</creator><general>Springer</general><general>Springer Nature B.V</general><scope>FBQ</scope><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>7SS</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</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>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>RC3</scope><scope>7QO</scope><scope>7X8</scope></search><sort><creationdate>20041101</creationdate><title>Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants</title><author>Park, J ; Lee, Y.K ; Kang, B.K ; Chung, W.I</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c411t-ed8edd9beb5ec3dbfd36ac7ccdd43e54dec3945e67c2abb99a15899b3a001ec63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>5-fluorocytosine</topic><topic>Agrobacterium tumefaciens</topic><topic>beta-glucuronidase</topic><topic>Biological and medical sciences</topic><topic>Blotting, Southern</topic><topic>Classical genetics, quantitative genetics, hybrids</topic><topic>codA gene</topic><topic>cytosine deaminase</topic><topic>DNA Primers</topic><topic>DNA, Bacterial - genetics</topic><topic>drug resistance</topic><topic>flucytosine</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes</topic><topic>Genetic Engineering - methods</topic><topic>genetic markers</topic><topic>Genetic Markers - genetics</topic><topic>genetic transformation</topic><topic>Genetic Vectors</topic><topic>Genetics of eukaryotes. Biological and molecular evolution</topic><topic>in vitro selection</topic><topic>negative selection</topic><topic>Nicotiana - genetics</topic><topic>Nicotiana tabacum</topic><topic>nptII gene</topic><topic>Plants, Genetically Modified</topic><topic>plasmid vectors</topic><topic>Polymerase Chain Reaction</topic><topic>Pteridophyta, spermatophyta</topic><topic>reporter genes</topic><topic>seed germination</topic><topic>tobacco</topic><topic>Transformation, Genetic - genetics</topic><topic>Transgenic plants</topic><topic>Vegetals</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Park, J</creatorcontrib><creatorcontrib>Lee, Y.K</creatorcontrib><creatorcontrib>Kang, B.K</creatorcontrib><creatorcontrib>Chung, W.I</creatorcontrib><collection>AGRIS</collection><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>Entomology Abstracts (Full archive)</collection><collection>Neurosciences Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</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>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>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>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>ProQuest Central China</collection><collection>Genetics Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Theoretical and applied genetics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Park, J</au><au>Lee, Y.K</au><au>Kang, B.K</au><au>Chung, W.I</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants</atitle><jtitle>Theoretical and applied genetics</jtitle><addtitle>Theor Appl Genet</addtitle><date>2004-11-01</date><risdate>2004</risdate><volume>109</volume><issue>8</issue><spage>1562</spage><epage>1567</epage><pages>1562-1567</pages><issn>0040-5752</issn><eissn>1432-2242</eissn><coden>THAGA6</coden><abstract>A negative selectable marker gene, codA, was successfully co-transformed with a GUS reporter gene to develop selectable marker gene-free transgenic plants. The pNC binary vector contained a T-DNA harboring the codA gene next to the nptII gene, while a second binary vector, pHG, contained a GUS reporter gene. Tobacco plants (Nicotiana tabacum cv. Samsun NN) were co-transformed via the mixture method with Agrobacterium tumefaciens LBA4404 strains harboring pNC and pHG, respectively. Seeds harvested from the co-transformants were sown on germination media containing 5-fluorocytosine (5-FC). Analysis of the progeny by GUS staining and PCR amplification revealed that all of the 5-FC-resistant R1 plants were codA free, and that the codA gene segregated independently of the GUS gene. Because codA-free seedlings developed normally on 5-FC-containing medium, we suggest that co-transformation with negatively selectable markers is a viable method for the production of easily distinguished, selectable marker gene-free transgenic plants.</abstract><cop>Heidelberg</cop><cop>Berlin</cop><pub>Springer</pub><pmid>15448898</pmid><doi>10.1007/s00122-004-1790-x</doi><tpages>6</tpages></addata></record> |
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| subjects | 5-fluorocytosine Agrobacterium tumefaciens beta-glucuronidase Biological and medical sciences Blotting, Southern Classical genetics, quantitative genetics, hybrids codA gene cytosine deaminase DNA Primers DNA, Bacterial - genetics drug resistance flucytosine Fundamental and applied biological sciences. Psychology Genes Genetic Engineering - methods genetic markers Genetic Markers - genetics genetic transformation Genetic Vectors Genetics of eukaryotes. Biological and molecular evolution in vitro selection negative selection Nicotiana - genetics Nicotiana tabacum nptII gene Plants, Genetically Modified plasmid vectors Polymerase Chain Reaction Pteridophyta, spermatophyta reporter genes seed germination tobacco Transformation, Genetic - genetics Transgenic plants Vegetals |
| title | Co-transformation using a negative selectable marker gene for the production of selectable marker gene-free transgenic plants |
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