Targeted integration and removal of transgenes in hybrid aspen (Populus tremula L. x P. tremuloides Michx.) using site-specific recombination systems.
Identifieur interne : 003097 ( Main/Exploration ); précédent : 003096; suivant : 003098Targeted integration and removal of transgenes in hybrid aspen (Populus tremula L. x P. tremuloides Michx.) using site-specific recombination systems.
Auteurs : M. Fladung [Allemagne] ; D. BeckerSource :
- Plant biology (Stuttgart, Germany) [ 1438-8677 ] ; 2010.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical : Integrases.
- genetics : Plants, Genetically Modified, Populus.
- Gene Transfer Techniques, Plasmids, Promoter Regions, Genetic, Recombination, Genetic, Transgenes.
Abstract
Two site-specific recombination systems, Cre/lox and FLP/FRT, were tested for marker gene removal and targeted gene transfer in a model tree system. A hybrid aspen clone (Populus tremula x Populus tremuloides) was co-transformed with plasmids containing either the FLP or the Cre recombinase, both under control of a heat-inducible promoter (HSP, Gmhsp17.5-E from soybean) flanked by the two recognition sites (FRT or lox). Molecular investigations of heat-shock treated Cre or FLP transgenic lines indicate excision of inserts between the two recognition sites. Further, a site-specific recombination at the FRT sites leading to targeted integration of a fragment could be demonstrated for the FLP/FRT system. Transgenic aspen carrying two constructs (each with different genes between the FRT sites) revealed (i) excision of both fragments between the FRT sites, and (ii) targeted integration of the fragment from the second construct exactly at the former position of the fragment in the first construct. These results indicate the usefulness of the two site-specific recombination systems in the tree species Populus. Combining both site-specific recombination systems, a strategy is suggested for targeted transgene transfer and removal of antibiotic marker genes.
DOI: 10.1111/j.1438-8677.2009.00293.x
PubMed: 20398239
Affiliations:
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Le document en format XML
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Fladung</name><affiliation wicri:level="1"><nlm:affiliation>Johann Heinrich von Thünen-Institut, Federal Research Institute for Rural Areas, Forestry and Fisheries, Institute of Forest Genetics, Grosshansdorf, Germany. matthias.fladung@vti.bund.de</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Johann Heinrich von Thünen-Institut, Federal Research Institute for Rural Areas, Forestry and Fisheries, Institute of Forest Genetics, Grosshansdorf</wicri:regionArea><wicri:noRegion>Grosshansdorf</wicri:noRegion><wicri:noRegion>Grosshansdorf</wicri:noRegion><wicri:noRegion>Grosshansdorf</wicri:noRegion></affiliation></author><author><name sortKey="Becker, D" sort="Becker, D" uniqKey="Becker D" first="D" last="Becker">D. Becker</name></author></analytic><series><title level="j">Plant biology (Stuttgart, Germany)</title><idno type="eISSN">1438-8677</idno><imprint><date when="2010" type="published">2010</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Gene Transfer Techniques (MeSH)</term><term>Integrases (MeSH)</term><term>Plants, Genetically Modified (genetics)</term><term>Plasmids (MeSH)</term><term>Populus (genetics)</term><term>Promoter Regions, Genetic (MeSH)</term><term>Recombination, Genetic (MeSH)</term><term>Transgenes (MeSH)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Integrases (MeSH)</term><term>Plasmides (MeSH)</term><term>Populus (génétique)</term><term>Recombinaison génétique (MeSH)</term><term>Régions promotrices (génétique) (MeSH)</term><term>Techniques de transfert de gènes (MeSH)</term><term>Transgènes (MeSH)</term><term>Végétaux génétiquement modifiés (génétique)</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Integrases</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Plants, Genetically Modified</term><term>Populus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Populus</term><term>Végétaux génétiquement modifiés</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Transfer Techniques</term><term>Plasmids</term><term>Promoter Regions, Genetic</term><term>Recombination, Genetic</term><term>Transgenes</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Integrases</term><term>Plasmides</term><term>Recombinaison génétique</term><term>Régions promotrices (génétique)</term><term>Techniques de transfert de gènes</term><term>Transgènes</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Two site-specific recombination systems, Cre/lox and FLP/FRT, were tested for marker gene removal and targeted gene transfer in a model tree system. A hybrid aspen clone (Populus tremula x Populus tremuloides) was co-transformed with plasmids containing either the FLP or the Cre recombinase, both under control of a heat-inducible promoter (HSP, Gmhsp17.5-E from soybean) flanked by the two recognition sites (FRT or lox). Molecular investigations of heat-shock treated Cre or FLP transgenic lines indicate excision of inserts between the two recognition sites. Further, a site-specific recombination at the FRT sites leading to targeted integration of a fragment could be demonstrated for the FLP/FRT system. Transgenic aspen carrying two constructs (each with different genes between the FRT sites) revealed (i) excision of both fragments between the FRT sites, and (ii) targeted integration of the fragment from the second construct exactly at the former position of the fragment in the first construct. These results indicate the usefulness of the two site-specific recombination systems in the tree species Populus. Combining both site-specific recombination systems, a strategy is suggested for targeted transgene transfer and removal of antibiotic marker genes.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">20398239</PMID><DateCompleted><Year>2010</Year><Month>06</Month><Day>28</Day></DateCompleted><DateRevised><Year>2010</Year><Month>12</Month><Day>28</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Electronic">1438-8677</ISSN><JournalIssue CitedMedium="Internet"><Volume>12</Volume><Issue>2</Issue><PubDate><Year>2010</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Plant biology (Stuttgart, Germany)</Title><ISOAbbreviation>Plant Biol (Stuttg)</ISOAbbreviation></Journal><ArticleTitle>Targeted integration and removal of transgenes in hybrid aspen (Populus tremula L. x P. tremuloides Michx.) using site-specific recombination systems.</ArticleTitle><Pagination><MedlinePgn>334-40</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1111/j.1438-8677.2009.00293.x</ELocationID><Abstract><AbstractText>Two site-specific recombination systems, Cre/lox and FLP/FRT, were tested for marker gene removal and targeted gene transfer in a model tree system. A hybrid aspen clone (Populus tremula x Populus tremuloides) was co-transformed with plasmids containing either the FLP or the Cre recombinase, both under control of a heat-inducible promoter (HSP, Gmhsp17.5-E from soybean) flanked by the two recognition sites (FRT or lox). Molecular investigations of heat-shock treated Cre or FLP transgenic lines indicate excision of inserts between the two recognition sites. Further, a site-specific recombination at the FRT sites leading to targeted integration of a fragment could be demonstrated for the FLP/FRT system. Transgenic aspen carrying two constructs (each with different genes between the FRT sites) revealed (i) excision of both fragments between the FRT sites, and (ii) targeted integration of the fragment from the second construct exactly at the former position of the fragment in the first construct. These results indicate the usefulness of the two site-specific recombination systems in the tree species Populus. Combining both site-specific recombination systems, a strategy is suggested for targeted transgene transfer and removal of antibiotic marker genes.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Fladung</LastName><ForeName>M</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Johann Heinrich von Thünen-Institut, Federal Research Institute for Rural Areas, Forestry and Fisheries, Institute of Forest Genetics, Grosshansdorf, Germany. matthias.fladung@vti.bund.de</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Becker</LastName><ForeName>D</ForeName><Initials>D</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType><PublicationType UI="D016454">Review</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Plant Biol (Stuttg)</MedlineTA><NlmUniqueID>101148926</NlmUniqueID><ISSNLinking>1435-8603</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>EC 2.7.7.-</RegistryNumber><NameOfSubstance UI="C045073">Cre recombinase</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 2.7.7.-</RegistryNumber><NameOfSubstance UI="D019426">Integrases</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><CommentsCorrectionsList><CommentsCorrections RefType="ErratumIn"><RefSource>Plant Biol (Stuttg). 2011 Jan;13(1):223</RefSource></CommentsCorrections></CommentsCorrectionsList><MeshHeadingList><MeshHeading><DescriptorName UI="D018014" MajorTopicYN="Y">Gene Transfer Techniques</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019426" MajorTopicYN="N">Integrases</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D030821" MajorTopicYN="N">Plants, Genetically Modified</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010957" MajorTopicYN="N">Plasmids</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D032107" MajorTopicYN="N">Populus</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011401" MajorTopicYN="N">Promoter Regions, Genetic</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011995" MajorTopicYN="N">Recombination, Genetic</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019076" MajorTopicYN="Y">Transgenes</DescriptorName></MeshHeading></MeshHeadingList><NumberOfReferences>44</NumberOfReferences></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2010</Year><Month>4</Month><Day>20</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2010</Year><Month>4</Month><Day>20</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2010</Year><Month>6</Month><Day>29</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">20398239</ArticleId><ArticleId IdType="pii">PLB293</ArticleId><ArticleId IdType="doi">10.1111/j.1438-8677.2009.00293.x</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Allemagne</li></country></list><tree><noCountry><name sortKey="Becker, D" sort="Becker, D" uniqKey="Becker D" first="D" last="Becker">D. Becker</name></noCountry><country name="Allemagne"><noRegion><name sortKey="Fladung, M" sort="Fladung, M" uniqKey="Fladung M" first="M" last="Fladung">M. Fladung</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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