Targeted mutagenesis in the progeny of maize transgenic plants |
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Authors: | Meizhu Yang Vesna Djukanovic Jessica Stagg Brian Lenderts Dennis Bidney S Carl Falco L Alexander Lyznik |
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Institution: | (1) Pioneer Hi-Bred International, A DuPont Business, Research Center, 7300 NW 62nd Avenue, Johnston, IA 50131-1004, USA;(2) Crop Genetics Research and Development, DuPont Agriculture & Nutrition, 7300 NW 62nd Avenue, Johnston, IA 50131-1004, USA |
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Abstract: | We have demonstrated that targeted mutagenesis can be accomplished in maize plants by excision, activation, and subsequent
elimination of an endonuclease in the progeny of genetic crosses. The yeast FLP/FRT site-specific recombination system was used to excise and transiently activate the previously integrated yeast I-SceI homing endonuclease in maize zygotes and/or developing embryos. An artificial I-SceI recognition sequence integrated into genomic DNA was analyzed for mutations to indicate the I-SceI endonuclease activity. Targeted mutagenesis of the I-SceI site occurred in about 1% of analyzed F1 plants. Short deletions centered on the I-SceI-produced double-strand break were the predominant genetic lesions observed in the F1 plants. The I-SceI expression cassette was not detected in the mutant F1 plants and their progeny. However, the original mutations were faithfully
transmitted to the next generation indicating that the mutations occurred early during the F1 plant development. The procedure
offers simultaneous production of double-strand breaks and delivery of DNA template combined with a large number of progeny
plants for future gene targeting experiments. |
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Keywords: | Double-strand break I-SceI Mutation Transgenic plant Maize Zea mays L |
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