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Introduction of pokeweed antiviral protein cDNA into Brassica napus and acquisition of transsenic plants resistant to viruses
Institution:ZHANG Haiyan , TIAN Yingchuan , ZHOU Yihua , DANG Benyuan LAN Haiyan , SONG Guiying , WANG Lanlan , LIU Guizhen , ZHANG Lihua and CHEN Zhenghua Institute of Genetics , Chinese Academy of Sciences , Beijing 100101, China; Institute of Microbiology , Chinese Academy of Sciences , Beijing 100080, China
Abstract:Using cotyledonary petioles as explants, the PAP cDNA controlled by wound-in- ducible promoter has been introduced into Brassica napus by coculture with Agrobacterium tumefaciens and laser microbeam puncture respectively. Regenerated plants resistant to gen-tamycin have been selected out. PCR amplification and Southern blotting analysis indicated that PAP cDNA together with wound-inducible promoter had been integrated into Brassica genome with transformation frequencies of 2.0% and 1.7% for two transformation methods respectively. The test of virus challenge showed that these transgenic Brassica plants were resistant in different degrees to mechanically inoculated TuMV.
Keywords:pokeweed antiviral protein cDNA  wound-inducible promoter  Agrobacterium tumefaciens  laser microbeam puncture  
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