甘蓝型油菜反义Fad2基因RNAi载体无选择标记转化研究

Research on marker-free transformation of Brassica napus with Fad2 antisense gene and RNA interference

利用甘蓝型油菜种子特异性贮存蛋白基因cruciferin启动子及NOS终止子构建了无选择标记基因且含有反义结构的油酸脱饱和酶基因(Fad2)的RNA i表达载体。通过农杆菌介导法,获得了无选择标记转基因植株;再生植株的PCR检测表明,外源基因已高频率地整合到油菜基因组中,而且获得的转基因种子通过脂肪酸含量分析表明,油酸含量比对照有了很大的提高。

A RNA interference expression cassette for Fad2 gene,which is marker-free gene and with antisense structure,was constructed u- sing the promotor of cruciferin gene from Brassica napus and NOS terminator. Transgenie rape plants without selection marker genes were ob- tained by Agrobacterium-mediated transformation. The results of PCR analysis indicated that Fad2 gene had been integrated into the genome of rapeseed in high efficiency. The fatty acid content analysis showed that the oleic acid content of transgenic seed was greatly increased as compared with control.