马铃薯sgt2基因启动子的克隆及其活性分析  

Cloning and GUS-aided activity analysis of potato sgt2 gene promoter

作　　者：魏桂民[1] 张金文[1] 王蒂[1] 张俊莲[1] 陆艳梅[1] 高宜峰[1] 

机构地区：[1]甘肃省作物遗传改良与种质创新重点实验室,甘肃农业大学农学院,甘肃兰州730070

出　　处：《甘肃农业大学学报》2014年第1期41-47,53,共8页Journal of Gansu Agricultural University

基　　金：国家自然科学基金项目(31260343);科技部国家重点基础研究发展计划(973计划)前期项目(2010CB13400);甘肃省自然科学基金项目(0710RJZA088)

摘　　要：糖苷生物碱(SGAs)是一类存在于茄科和百合科植物的重要次生代谢物,与植物的抗逆性和产品品质有密切关系.茄啶葡糖基转移酶(SGT2)是SGAs合成代谢途径的末端关键酶之一,研究其编码基因的启动子序列对于SGAs生物合成代谢调控有重要的作用和意义.本研究采用染色体步移技术,克隆到马铃薯茄啶葡糖基转移酶基因(sgt2)起始密码子上游2 098bp的启动子序列,已注册到GenBank(注册号:KC331038).构建该启动子驱动报告基因gfp∶∶gus的植物双元表达载体p1304sgt2p,采用农杆菌介导的烟草叶片瞬时表达,通过GUS组织化学染色分析sgt2p启动子的活性.结果表明:gus基因在转化烟草叶片中高效表达,克隆的启动子具有活性.The potato steroidal glycoalkaloids（SGAs） is a kind of important secondary metabolite in So- lanace and Liliaceae. It is closely related to the stress resistance and product quality of plant. Solanidine glu- cosyltransferase is one of the terminases of SGAs synthetic metabolic pathway,to study the promoter of its coding gene has important significance in the regulation of the SGAs biosynthesis metabolism in plant. A 2 098 bp promoter sequence of 5~ upstream of initial codon of sgt2 gene was obtained from Solanum tubero- sum by using the genome walking. The promoter sequence had been submitted to the GenBank（accession number. KC331038）. The promoter sequence was fused with gfp：＂ gus gene originated from pCAM- BIA1304 to construct a binary vector p1304sgt2p, which was introduced into wild type tobacco leaves through Arobacterium-mediated transformation. The sgt2 gene promoter activity in sgt2p：gfp：gus transgenic tobacco leaves was determined through histochemical staining. The GUS staining showed that gus gene was highly expressed in transgenic tobacco leaves, the promoter cloned had activity.

关 键 词：马铃薯 糖苷生物碱 茄啶葡糖基转移酶 启动子克隆 瞬时表达 

分 类 号：S532[农业科学—作物学]