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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pubmed:16593993Le document en format XML
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<affiliation><nlm:affiliation>Plant Gene Expression Center, U.S. Department of Agriculture, Agricultural Research Service, 800 Buchanan Street, Albany, CA 94710.</nlm:affiliation>
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<author><name sortKey="Harper, E C" sort="Harper, E C" uniqKey="Harper E" first="E C" last="Harper">E C Harper</name>
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<author><name sortKey="Svab, Z" sort="Svab, Z" uniqKey="Svab Z" first="Z" last="Svab">Z. Svab</name>
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<author><name sortKey="Sanford, J C" sort="Sanford, J C" uniqKey="Sanford J" first="J C" last="Sanford">J C Sanford</name>
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<author><name sortKey="Fromm, M E" sort="Fromm, M E" uniqKey="Fromm M" first="M E" last="Fromm">M E Fromm</name>
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<author><name sortKey="Maliga, P" sort="Maliga, P" uniqKey="Maliga P" first="P" last="Maliga">P. Maliga</name>
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<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>
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<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. 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.</AbstractText>
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