Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2013, Vol. 29 ›› Issue (21): 131-136.doi: 10.11924/j.issn.1000-6850.2012-3125

Special Issue: 生物技术 油料作物

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Construction of Soybean FAD2-1b Gene Silencing Vector

  

  • Received:2012-09-13 Revised:2012-10-25 Online:2013-07-25 Published:2013-07-25

Abstract: The soybean FAD2-1b gene was amplified by RT-PCR from transgenic soybeans, and soybean FAD2-1b gene silencing vector was constructed for breeding transgenic soybeans with biosafety. Seed specific promoter GY1 of soybean was amplified from high protein soybean ‘Heinong 35 ’; FAD2-1b gene introns 1 was amplified from high oil soybean ‘Heinong 37’; FAD2-1b gene fragment was amplified from five transgenic acceptors soybean, it was used as forward arm and reverse arm. These target fragments were ligated to construct a double T-DNA with bar gene selection marker, promoter GY1 and FAD2-1b gene intron 1. Then double T-DNA was introduced into E. coli DH5α. The cloning of recombination plasmids was identified correctly by enzymes digestion and sequence analysis. The recombination plasmid was named pDT-FAD2I. FAD2-1b gene silencing vector has been constructed. This will constitute an important foundation for further development of FAD2-1b gene silencing soybeans and high quality soybean transgenic security.

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