绿豆种子8S球蛋白基因启动子的克隆及分析  

Molecular Cloning and Characterization of the Promoter of Vigna radiata 8S Globulin Gene

作　　者：徐超[1] 杨悦宁[1] 王吟[1] 张梦晗[1] 谢伟红[1] 杨维东[1] 刘洁生[1] 李宏业[1] 

机构地区：[1]暨南大学生物工程学系,广州510632

出　　处：《西北植物学报》2011年第2期242-247,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：广东省自然科学基金(9151063101000001);教育部高等学校博士学科点专项科研基金(20094401120010);国家大学生创新性实验计划基金(081055908)

摘　　要：根据绿豆种子8S球蛋白α′亚基基因的末端序列设计3个特异反向引物.以绿豆基因组DNA为模板,采用基因组步移法,获得了8S球蛋白α′亚基基因起始密码子上游784 bp的DNA片段,通过序列测定和生物信息学分析,发现该序列含有启动子核心区以及大量的种子特异表达相关的顺式作用元件,表明此序列为种子特异启动子序列.通过PCR方法在启动子3′端加上翻译增强序列TMV-omega序列,构建了植物双元表达载体pBI-8SGα-′o-mega-gus,并成功将其转化进入农杆菌.According to the sequence of Vigna radiata 8S globulin α′ isoform gene from GenBank,we designed and synthesized three reverse primers.A DNA fragment of 784 bp up stream of the coding sequence of V.radiata 8S globulin α′ isoform gene was amplified by Genome Walking with the genomic DNA of mung bean as the template.Sequence analysis suggested that the fragment is the seed specific promoter.Subjoined the TMV-omega sequence in the 3′ end of promoter of V.radiata 8S globulin alpha isoform gene by PCR.Constructed plant binary expression vector pBI-8SG α′-omega-gus,and then transformed into Agrobacterium cells.

关 键 词：绿豆8S球蛋白 种子特异启动子 基因组步移 

分 类 号：Q785[生物学—分子生物学]