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

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.