编码拟南芥含BAG结构域蛋白的原核表达载体构建

生物信息学分析显示,拟南芥基因At5g62390编码一种钙调素结合蛋白,在其钙调素结合结构域有一个BAG(Bcl-2-associated athnogene)结构域存在,与钙调素结构域部分重叠.为了从实验上进一步研究该蛋白的钙调素结合特征及BAG结构域在钙调素结合中可能的调节作用,通过聚合酶链式反应(PCR)扩增不同结构域编码区的cDNA序列,构建到原核表达载体pET32-a中.测序分析表明:目的序列已正确克隆到表达载体上,为进一步表达目的蛋白及其不同结构域用于生化功能鉴定打下了基础.

Construction of the recombinant prokaryotic expression plasmid contain BAG domain in Arabidopsis thaliana

Bioinformatics analysis of Arabidopsis gene At5g62390 encoded a calmodulin-binding protein, there was a BAG domain in its calcium-binding domain and the calmodulin-domain overlapped. To further experimental studied the characteristics of the protein calmodulin-binding and the modulation of BAG domain in calmodulin-binding. The coding domain of different cDNAs were amplified by PCR (polymerase chain reaction), and subcloned into prokaryotic expression vector pET32-a. DNA sequencing showed that the sequences had been correctly inserted into the recombinant plasmids. This work was a good start for further study the expression of the target proteins and identification the biochemical functions of different domains.