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Genetic transformation of Nicotiana africana Merxm. with plasmids containing lox recombination sites

Shcherbak N.L., Belokurova V.B., Komarnitski I.K., Kuchuk N.V.

 


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An efficient genetic transformation method for african tobacco Nicotiana africana Merxm. has been established. African tobacco is a valuable source for cytoplasmic male sterility (CMS) and nuclear encoded resistance to potato virus Y (PVY). N. africana transgenic plants have been obtained using both Agrobacterium-mediated and direct transformation of leaf explants with gold particle bombardment using particle inflow gun. Plasmid vectors containing phosphinothricin resistance gene (bar gene) coding region without promoter and independent 35S promoter between lox sites (lox-bar-35S-lox) and nptII gene were used. Transgenic plants were selected according to growth capacity on the selective medium containing 50 mg/l kanamycin. PCR analyses of kanamycin-resistant plants confirmed the presence of nptII and bar genes in their genome. Agrobacterium-mediated transformation of root explants has proved to be the most efficient transformation method for N. africana.

Tsitologiya i Genetika 2004, vol. 38, no. 4, pp. 3-8



Shcherbak N.L., Belokurova V.B., Komarnitski I.K., Kuchuk N.V. Genetic transformation of Nicotiana africana Merxm. with plasmids containing lox recombination sites, Tsitol Genet., 2004, vol. 38, no. 4, pp. 3-8.




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