普通烟草IRL基因反义植物表达载体的构建  

Construction of an Antisense Plant Expression Vector of IRL from Common Tobacco

作　　者：陈伟 马建光 李红勋 潘文杰 

机构地区：[1]贵州省烟草科学研究院,贵阳550081 [2]贵州省威宁县烟草分公司,贵州威宁553100

出　　处：《湖北农业科学》2014年第19期4730-4733,共4页Hubei Agricultural Sciences

基　　金：贵州省科学技术基金项目[黔科合J字(2011)2340号];国家烟草专卖局科技重大专项(Ts-02-20110015)

摘　　要：异黄酮还原酶相似蛋白(IRL)是与异黄酮还原酶具有高度序列同源一致性而功能不同的一类蛋白。通过PCR扩增普通烟草(Nicotiana tabacum)品种龙里红花烟IRL基因的一段特异保守片段Nt IRLA,并将其亚克隆到中间载体p CAMBIA2301G中,构建了IRL反义植物表达载体p CAMBIA2301G-IRLA。通过PCR鉴定、酶切鉴定证实载体构建成功,并转化到根癌农杆菌LBA4404菌株中形成工程菌株,为进一步利用转基因手段明确IRL基因在烟草次生代谢中的功能奠定了基础。Isoflavone reductase-like protein has highly homologous consistency of sequences with isoflavone reductase,but their function was different.In order to reveal its function in metabolism pathway,a specific conserved fragment of Nt IRLA from Nicotiana tabacum Longlihonghua was amplified by PCR and subcloned into intermediate vector p CAMBIA2301 G.An antisense plant expression vector p CAMBIA2301G-IRLA was constructed,confirmed by PCR analysis and endonuclease digestion.Then the recombinant vector was introduced into agrobacterium tumefaciens strain LBA4404 to form an engineer ed strain.It will lay a foundation for studying biological function of IRL gene involved in tobacco metabolism with transgenic approach.

关 键 词：类异黄酮还原酶基因 反义表达载体 普通烟草 

分 类 号：S572[农业科学—烟草工业]