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Gene Targeting by Homologous Recombination as a Biotechnological Tool for Rice Functional Genomics
The modification of an endogenous gene into a designed sequence by homologous recombination, termed gene targeting (GT), has broad implications for basic and applied research. Rice (Oryza sativa), with a sequenced genome of 389 Mb, is one of the most important crops and a model plant for cereals, an...
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| Published in: | Plant physiology (Bethesda) 2007-06, Vol.144 (2), p.846-856 |
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| Main Authors: | , , , , |
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| cites | cdi_FETCH-LOGICAL-c539t-360d5fe0dd2d9dfaf426582e49aa07cecdbce4cb924c984a9314aa69322fe8f93 |
| container_end_page | 856 |
| container_issue | 2 |
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| container_title | Plant physiology (Bethesda) |
| container_volume | 144 |
| creator | Terada, Rie Johzuka-Hisatomi, Yasuyo Saitoh, Miho Asao, Hisayo Iida, Shigeru |
| description | The modification of an endogenous gene into a designed sequence by homologous recombination, termed gene targeting (GT), has broad implications for basic and applied research. Rice (Oryza sativa), with a sequenced genome of 389 Mb, is one of the most important crops and a model plant for cereals, and the single-copy gene Waxy on chromosome 6 has been modified with a frequency of 1% per surviving callus by GT using a strong positive-negative selection. Because the strategy is independent of gene-specific selection or screening, it is in principle applicable to any gene. However, a gene in the multigene family or a gene carrying repetitive sequences may preclude efficient homologous recombination-promoted GT due to the occurrence of ectopic recombination. Here, we describe an improved GT procedure whereby we obtained nine independent transformed calli having the alcohol dehydrogenase2 (Adh2) gene modified with a frequency of approximately 2% per surviving callus and subsequently isolated eight fertile transgenic plants without the concomitant occurrence of undesirable ectopic events, even though the rice genome carries four Adh genes, including a newly characterized Adh3 gene, and a copy of highly repetitive retroelements is present adjacent to the Adh2 gene. The results indicate that GT using a strong positive-negative selection can be widely applicable to functional genomics in rice and presumably in other higher plants. |
| doi_str_mv | 10.1104/pp.107.095992 |
| format | article |
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Rice (Oryza sativa), with a sequenced genome of 389 Mb, is one of the most important crops and a model plant for cereals, and the single-copy gene Waxy on chromosome 6 has been modified with a frequency of 1% per surviving callus by GT using a strong positive-negative selection. Because the strategy is independent of gene-specific selection or screening, it is in principle applicable to any gene. However, a gene in the multigene family or a gene carrying repetitive sequences may preclude efficient homologous recombination-promoted GT due to the occurrence of ectopic recombination. Here, we describe an improved GT procedure whereby we obtained nine independent transformed calli having the alcohol dehydrogenase2 (Adh2) gene modified with a frequency of approximately 2% per surviving callus and subsequently isolated eight fertile transgenic plants without the concomitant occurrence of undesirable ectopic events, even though the rice genome carries four Adh genes, including a newly characterized Adh3 gene, and a copy of highly repetitive retroelements is present adjacent to the Adh2 gene. The results indicate that GT using a strong positive-negative selection can be widely applicable to functional genomics in rice and presumably in other higher plants.</description><identifier>ISSN: 0032-0889</identifier><identifier>ISSN: 1532-2548</identifier><identifier>EISSN: 1532-2548</identifier><identifier>DOI: 10.1104/pp.107.095992</identifier><identifier>PMID: 17449652</identifier><identifier>CODEN: PPHYA5</identifier><language>eng</language><publisher>Rockville, MD: American Society of Plant Biologists</publisher><subject>Alcohol Dehydrogenase - genetics ; Biological and medical sciences ; Breakthrough Technologies ; Bryopsida - genetics ; Callus ; Fundamental and applied biological sciences. Psychology ; Gene targeting ; Gene Targeting - methods ; Genes ; Genes. Genome ; Genetic vectors ; Genomes ; Genomics - methods ; Molecular and cellular biology ; Molecular genetics ; Molecular Sequence Data ; Mosses ; Oryza - genetics ; Oryza sativa ; Plants ; Plants, Genetically Modified ; Polymerase chain reaction ; Recombination, Genetic ; Rice ; Transgenes</subject><ispartof>Plant physiology (Bethesda), 2007-06, Vol.144 (2), p.846-856</ispartof><rights>Copyright 2007 American Society of Plant Biologists</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c539t-360d5fe0dd2d9dfaf426582e49aa07cecdbce4cb924c984a9314aa69322fe8f93</citedby><cites>FETCH-LOGICAL-c539t-360d5fe0dd2d9dfaf426582e49aa07cecdbce4cb924c984a9314aa69322fe8f93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/40065534$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/40065534$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>315,786,790,27957,27958,58593,58826</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18846713$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17449652$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Terada, Rie</creatorcontrib><creatorcontrib>Johzuka-Hisatomi, Yasuyo</creatorcontrib><creatorcontrib>Saitoh, Miho</creatorcontrib><creatorcontrib>Asao, Hisayo</creatorcontrib><creatorcontrib>Iida, Shigeru</creatorcontrib><title>Gene Targeting by Homologous Recombination as a Biotechnological Tool for Rice Functional Genomics</title><title>Plant physiology (Bethesda)</title><addtitle>Plant Physiol</addtitle><description>The modification of an endogenous gene into a designed sequence by homologous recombination, termed gene targeting (GT), has broad implications for basic and applied research. 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Here, we describe an improved GT procedure whereby we obtained nine independent transformed calli having the alcohol dehydrogenase2 (Adh2) gene modified with a frequency of approximately 2% per surviving callus and subsequently isolated eight fertile transgenic plants without the concomitant occurrence of undesirable ectopic events, even though the rice genome carries four Adh genes, including a newly characterized Adh3 gene, and a copy of highly repetitive retroelements is present adjacent to the Adh2 gene. The results indicate that GT using a strong positive-negative selection can be widely applicable to functional genomics in rice and presumably in other higher plants.</description><subject>Alcohol Dehydrogenase - genetics</subject><subject>Biological and medical sciences</subject><subject>Breakthrough Technologies</subject><subject>Bryopsida - genetics</subject><subject>Callus</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene targeting</subject><subject>Gene Targeting - methods</subject><subject>Genes</subject><subject>Genes. Genome</subject><subject>Genetic vectors</subject><subject>Genomes</subject><subject>Genomics - methods</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Mosses</subject><subject>Oryza - genetics</subject><subject>Oryza sativa</subject><subject>Plants</subject><subject>Plants, Genetically Modified</subject><subject>Polymerase chain reaction</subject><subject>Recombination, Genetic</subject><subject>Rice</subject><subject>Transgenes</subject><issn>0032-0889</issn><issn>1532-2548</issn><issn>1532-2548</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><recordid>eNqF0U1v1DAQBmALgei2cOQI-AK3LOOvxD6WirZIlZDK9hxNHHtxlcTBzh767_Eqq_bIySPNo1ea14R8YLBlDOS3ed4yaLZglDH8FdkwJXjFldSvyQagzKC1OSPnOT8CABNMviVnrJHS1IpvSHfjJkd3mPZuCdOedk_0No5xiPt4yPTe2Th2YcIlxIlipki_h7g4-2c6kmBxoLsYB-pjovfBOnp9mOwRl0VJjmOw-R1543HI7v3pvSAP1z92V7fV3a-bn1eXd5VVwiyVqKFX3kHf8970Hr3ktdLcSYMIjXW276yTtjNcWqMlmnIKYm0E595pb8QF-brmzin-Pbi8tGPI1g0DTq4c0zZQQ1Nr9V_ITF06U7zAaoU2xZyT8-2cwojpqWXQHttv57mMTbu2X_ynU_ChG13_ok91F_DlBDCX7nzCyYb84rSWdcNEcR9X95iXmJ73EqBWSsiy_7zuPcYW96lkPPzm5XcBGs00A_EPw-Ogkg</recordid><startdate>20070601</startdate><enddate>20070601</enddate><creator>Terada, Rie</creator><creator>Johzuka-Hisatomi, Yasuyo</creator><creator>Saitoh, Miho</creator><creator>Asao, Hisayo</creator><creator>Iida, Shigeru</creator><general>American Society of Plant Biologists</general><general>American Society of Plant Physiologists</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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20070601</creationdate><title>Gene Targeting by Homologous Recombination as a Biotechnological Tool for Rice Functional Genomics</title><author>Terada, Rie ; Johzuka-Hisatomi, Yasuyo ; Saitoh, Miho ; Asao, Hisayo ; Iida, Shigeru</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c539t-360d5fe0dd2d9dfaf426582e49aa07cecdbce4cb924c984a9314aa69322fe8f93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Alcohol Dehydrogenase - genetics</topic><topic>Biological and medical sciences</topic><topic>Breakthrough Technologies</topic><topic>Bryopsida - genetics</topic><topic>Callus</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene targeting</topic><topic>Gene Targeting - methods</topic><topic>Genes</topic><topic>Genes. Genome</topic><topic>Genetic vectors</topic><topic>Genomes</topic><topic>Genomics - methods</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Mosses</topic><topic>Oryza - genetics</topic><topic>Oryza sativa</topic><topic>Plants</topic><topic>Plants, Genetically Modified</topic><topic>Polymerase chain reaction</topic><topic>Recombination, Genetic</topic><topic>Rice</topic><topic>Transgenes</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Terada, Rie</creatorcontrib><creatorcontrib>Johzuka-Hisatomi, Yasuyo</creatorcontrib><creatorcontrib>Saitoh, Miho</creatorcontrib><creatorcontrib>Asao, Hisayo</creatorcontrib><creatorcontrib>Iida, Shigeru</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>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Plant physiology (Bethesda)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Terada, Rie</au><au>Johzuka-Hisatomi, Yasuyo</au><au>Saitoh, Miho</au><au>Asao, Hisayo</au><au>Iida, Shigeru</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Gene Targeting by Homologous Recombination as a Biotechnological Tool for Rice Functional Genomics</atitle><jtitle>Plant physiology (Bethesda)</jtitle><addtitle>Plant Physiol</addtitle><date>2007-06-01</date><risdate>2007</risdate><volume>144</volume><issue>2</issue><spage>846</spage><epage>856</epage><pages>846-856</pages><issn>0032-0889</issn><issn>1532-2548</issn><eissn>1532-2548</eissn><coden>PPHYA5</coden><notes>http://www.plantphysiol.org/</notes><notes>ObjectType-Article-2</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-1</notes><notes>content type line 23</notes><notes>ObjectType-Article-1</notes><notes>ObjectType-Feature-2</notes><abstract>The modification of an endogenous gene into a designed sequence by homologous recombination, termed gene targeting (GT), has broad implications for basic and applied research. Rice (Oryza sativa), with a sequenced genome of 389 Mb, is one of the most important crops and a model plant for cereals, and the single-copy gene Waxy on chromosome 6 has been modified with a frequency of 1% per surviving callus by GT using a strong positive-negative selection. Because the strategy is independent of gene-specific selection or screening, it is in principle applicable to any gene. However, a gene in the multigene family or a gene carrying repetitive sequences may preclude efficient homologous recombination-promoted GT due to the occurrence of ectopic recombination. Here, we describe an improved GT procedure whereby we obtained nine independent transformed calli having the alcohol dehydrogenase2 (Adh2) gene modified with a frequency of approximately 2% per surviving callus and subsequently isolated eight fertile transgenic plants without the concomitant occurrence of undesirable ectopic events, even though the rice genome carries four Adh genes, including a newly characterized Adh3 gene, and a copy of highly repetitive retroelements is present adjacent to the Adh2 gene. The results indicate that GT using a strong positive-negative selection can be widely applicable to functional genomics in rice and presumably in other higher plants.</abstract><cop>Rockville, MD</cop><pub>American Society of Plant Biologists</pub><pmid>17449652</pmid><doi>10.1104/pp.107.095992</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0032-0889 |
| ispartof | Plant physiology (Bethesda), 2007-06, Vol.144 (2), p.846-856 |
| issn | 0032-0889 1532-2548 1532-2548 |
| language | eng |
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| source | Oxford University Press Journals; JSTOR Archival Journals and Primary Sources Collection |
| subjects | Alcohol Dehydrogenase - genetics Biological and medical sciences Breakthrough Technologies Bryopsida - genetics Callus Fundamental and applied biological sciences. Psychology Gene targeting Gene Targeting - methods Genes Genes. Genome Genetic vectors Genomes Genomics - methods Molecular and cellular biology Molecular genetics Molecular Sequence Data Mosses Oryza - genetics Oryza sativa Plants Plants, Genetically Modified Polymerase chain reaction Recombination, Genetic Rice Transgenes |
| title | Gene Targeting by Homologous Recombination as a Biotechnological Tool for Rice Functional Genomics |
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