Stable genetic transformation of intact Nicotiana cells by the particle bombardment process.
Identifieur interne : 000965 ( Main/Corpus ); précédent : 000964; suivant : 000966Stable genetic transformation of intact Nicotiana cells by the particle bombardment process.
Auteurs : T M Klein ; E C Harper ; Z. Svab ; J C Sanford ; M E Fromm ; P. MaligaSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 1988.
Abstract
We show that the genetic transformation of Nicotiana tabacum can be achieved by bombarding intact cells and tissues with DNA-coated particles. Leaves or suspension culture cells were treated with tungsten microprojectiles carrying plasmid DNA containing a neomycin phosphotransferase gene. Callus harboring the foreign gene was recovered from the bombarded tissue by selection on medium containing kanamycin. Kanamycin-resistant plants have subsequently been regenerated from the callus derived from leaves. Transient expression of an introduced beta-glucuronidase gene was used to assess the efficiency of DNA delivery by microprojectiles. The frequency of cells that were stably transformed with the neomycin phosphotransferase gene was a few percent of the cells that transiently expressed the beta-glucuronidase gene. These results show that gene transfer by high-velocity microprojectiles is a rapid and direct means for transforming intact plant cells and tissues that eliminates the need for production of protoplasts or infection by Agrobacterium.
DOI: 10.1073/pnas.85.22.8502
PubMed: 16593993
PubMed Central: PMC282486
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Svab</name></author><author><name sortKey="Sanford, J C" sort="Sanford, J C" uniqKey="Sanford J" first="J C" last="Sanford">J C Sanford</name></author><author><name sortKey="Fromm, M E" sort="Fromm, M E" uniqKey="Fromm M" first="M E" last="Fromm">M E Fromm</name></author><author><name sortKey="Maliga, P" sort="Maliga, P" uniqKey="Maliga P" first="P" last="Maliga">P. 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Svab</name></author><author><name sortKey="Sanford, J C" sort="Sanford, J C" uniqKey="Sanford J" first="J C" last="Sanford">J C Sanford</name></author><author><name sortKey="Fromm, M E" sort="Fromm, M E" uniqKey="Fromm M" first="M E" last="Fromm">M E Fromm</name></author><author><name sortKey="Maliga, P" sort="Maliga, P" uniqKey="Maliga P" first="P" last="Maliga">P. Maliga</name></author></analytic><series><title level="j">Proceedings of the National Academy of Sciences of the United States of America</title><idno type="ISSN">0027-8424</idno><imprint><date when="1988" type="published">1988</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We show that the genetic transformation of Nicotiana tabacum can be achieved by bombarding intact cells and tissues with DNA-coated particles. Leaves or suspension culture cells were treated with tungsten microprojectiles carrying plasmid DNA containing a neomycin phosphotransferase gene. Callus harboring the foreign gene was recovered from the bombarded tissue by selection on medium containing kanamycin. Kanamycin-resistant plants have subsequently been regenerated from the callus derived from leaves. Transient expression of an introduced beta-glucuronidase gene was used to assess the efficiency of DNA delivery by microprojectiles. The frequency of cells that were stably transformed with the neomycin phosphotransferase gene was a few percent of the cells that transiently expressed the beta-glucuronidase gene. These results show that gene transfer by high-velocity microprojectiles is a rapid and direct means for transforming intact plant cells and tissues that eliminates the need for production of protoplasts or infection by Agrobacterium.</div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">16593993</PMID><DateCompleted><Year>2010</Year><Month>06</Month><Day>30</Day></DateCompleted><DateRevised><Year>2019</Year><Month>05</Month><Day>01</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0027-8424</ISSN><JournalIssue CitedMedium="Print"><Volume>85</Volume><Issue>22</Issue><PubDate><Year>1988</Year><Month>Nov</Month></PubDate></JournalIssue><Title>Proceedings of the National Academy of Sciences of the United States of America</Title><ISOAbbreviation>Proc Natl Acad Sci U S A</ISOAbbreviation></Journal><ArticleTitle>Stable genetic transformation of intact Nicotiana cells by the particle bombardment process.</ArticleTitle><Pagination><MedlinePgn>8502-5</MedlinePgn></Pagination><Abstract><AbstractText>We show that the genetic transformation of Nicotiana tabacum can be achieved by bombarding intact cells and tissues with DNA-coated particles. Leaves or suspension culture cells were treated with tungsten microprojectiles carrying plasmid DNA containing a neomycin phosphotransferase gene. Callus harboring the foreign gene was recovered from the bombarded tissue by selection on medium containing kanamycin. Kanamycin-resistant plants have subsequently been regenerated from the callus derived from leaves. Transient expression of an introduced beta-glucuronidase gene was used to assess the efficiency of DNA delivery by microprojectiles. The frequency of cells that were stably transformed with the neomycin phosphotransferase gene was a few percent of the cells that transiently expressed the beta-glucuronidase gene. 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