甘蓝型油菜和Lesquerella fendleri fad2基因片段克隆及hpRNA植物表达载体的构建  

Cloning and hpRNA vector construction for fad2 genes in Brassica napus and Lesquerella fendleri

作　　者：汪承刚[1] 王文燕[2] 谢妤[1] 黄邦全[1] 

机构地区：[1]湖北大学生命科学学院,湖北武汉430062 [2]武汉生物工程学院,湖北武汉430415

出　　处：《湖北大学学报（自然科学版）》2007年第1期89-92,共4页Journal of Hubei University：Natural Science

基　　金：湖北省科技厅国际合作重点项目(2005CA020);湖北省教育厅国际合作项目(G200510001)资助

摘　　要：首先利用PCR从甘蓝型油菜(Brassica napus L.)和Lesquerella fendleri基因组DNA中分别扩增出Δ12-脂肪酸去饱和酶fad2基因片段,再以扩增出的片段为模板设计引物从一端扩增出相应的小片段,然后将同一基因的大小片段反向连接,插入到种子特异表达载体2300-nap多克隆位点的napin启动子和nos终止子之间,构建成可以在种子中转录表达发夹RNA(Hairpin RNA,hpRNA)结构的植物表达载体.Delta-12 desaurase （FAD2） plays an important role in the poly-unsaturated fatty acid synthesis. Gene fragments for fad2 was PCR-amplified from B. napus and L. fendleri genomic DNA and used as templates to amplify a smaller fragment. The larger and the smaller fragments of the same gene were ligated in opposite orientations into a seed specific expression vector 2300-nap to produce hpRNA vectors. Sequence analyses indicated that the fragments we cloned were indeed genes for fad2 in B. napus and L. fendleri.

关 键 词：甘蓝型油菜 Lesquerella fendleri FAD2基因 发夹RNA(hpRNA) 载体构建 

分 类 号：Q78[生物学—分子生物学] S565.403.2[农业科学—作物学]