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Current Genetics
Yehuda, H., Department of Molecular Genetics, MIGAL Galilee Technological Center, P.O. Box 90000, Rosh Pina 12100, Israel
Droby, S., Department of Postharvest Science, Agricultural Research Organization, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Wisniewski, M., Appalachian Fruit Research Station, USDA-ARS, 45 Wiltshire Road, Kearneysville, WV 25430, United States
Goldway, M., Department of Molecular Genetics, MIGAL Galilee Technological Center, P.O. Box 90000, Rosh Pina 12100, Israel
Lithium acetate transformation and electroporation were applied to the biocontrol yeast, Candida oleophila. The hygromycin B resistance gene, flanked by the phosphoglycerate kinase promoter and terminator of Candida tropicalis, served as the genetic selection marker. The transformation efficiency of electroporation was almost 400 times more efficient than that of the lithium acetate method. While incorporation of DNA, flanked by a sequence endogenous to C. oleophila, transpired apparently by homologous recombination, the integration of DNA (that did not contain C. oleophila DNA) occurred at random. Whereas transformants were observed with a linear segment of the plasmid, none were detected with the undigested plasmid. This system provides both a tool for the molecular analysis of the biocontrol mechanism of C. oleophila and a means of tagging C. oleophila for field studies.
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A transformation system for the biocontrol yeast, Candida oleophila, based on hygromycin B resistance
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Yehuda, H., Department of Molecular Genetics, MIGAL Galilee Technological Center, P.O. Box 90000, Rosh Pina 12100, Israel
Droby, S., Department of Postharvest Science, Agricultural Research Organization, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel
Wisniewski, M., Appalachian Fruit Research Station, USDA-ARS, 45 Wiltshire Road, Kearneysville, WV 25430, United States
Goldway, M., Department of Molecular Genetics, MIGAL Galilee Technological Center, P.O. Box 90000, Rosh Pina 12100, Israel
A transformation system for the biocontrol yeast, Candida oleophila, based on hygromycin B resistance
Lithium acetate transformation and electroporation were applied to the biocontrol yeast, Candida oleophila. The hygromycin B resistance gene, flanked by the phosphoglycerate kinase promoter and terminator of Candida tropicalis, served as the genetic selection marker. The transformation efficiency of electroporation was almost 400 times more efficient than that of the lithium acetate method. While incorporation of DNA, flanked by a sequence endogenous to C. oleophila, transpired apparently by homologous recombination, the integration of DNA (that did not contain C. oleophila DNA) occurred at random. Whereas transformants were observed with a linear segment of the plasmid, none were detected with the undigested plasmid. This system provides both a tool for the molecular analysis of the biocontrol mechanism of C. oleophila and a means of tagging C. oleophila for field studies.
Scientific Publication
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