甘蓝型油菜乙醇酸氧化酶(GO)基因片段的克隆及hpRNA表达载体构建  

Cloning and hpRNA vector construction for Brassica napus glycolate oxidase gene(GO)

作　　者：汪承刚[1] 李长春[1] 谢妤[1] 刘幼琪[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

出　　处：《湖北大学学报（自然科学版）》2007年第3期286-289,324,共5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：通过PCR从甘蓝型油菜(Brassica napus)华双4号基因组DNA中扩增出乙醇酸氧化酶(GO)基因片段,DNA序列分析表明扩增GO基因片段的外显子部分与报道序列相同.以扩增出的GO基因片段作模板从一端设计引物扩增出一个相应的小片段.将GO基因大小两个片段反向连接,插入到植物表达载体2300-nap的napin启动子和nos终止子之间,植物表达载体2300-35S的35S启动子和nos终止子之间,分别构建成可转录表达出发夹RNA(hairpin RNA,hpRNA)结构的种子特异型和组成型油菜hpRNA干扰载体.Glycolate oxidase （GO） is a key enzyme for plant photorespiration. GO Gene fragment was PCR-amplified from B. napus genomic DNA and then used as template to amplify a smaller fragment. The larger and the smaller fragments were ligated in opposite orientations into a seed specific expression vector 2300-nap and constitutive vector 2300-35S respectively to produce hpRNAi vector hp-GO-nap and hp-GO-35S. Sequence analysis indicated that the GO fragment we amplified had 100% similarity in the exon part with the published sequence in the Genbank.

关 键 词：甘蓝型油菜 乙醇酸氧化酶(GO) 光呼吸 净光合速率 发夹RNA(hpRNA) 载体构建 

分 类 号：Q785[生物学—分子生物学] S565.403.2[农业科学—作物学]