小麦甲基结合蛋白基因MBD3重组表达载体的构建和原核表达  

Recombinant Expression Vector with Wheat MBD3 Gene and its Prokaryotic Expression

作　　者：孟凡荣[1] 司志飞[1] 刘昊英[1] 张问[1] 

机构地区：[1]河南农业大学生命科学学院,郑州450002

出　　处：《中国农学通报》2008年第11期78-80,共3页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金项目"小麦DNA甲基化时空差异及甲基结合蛋白基因的克隆和表达"(30300195)。

摘　　要：DNA甲基化在基因表达调控过程中发挥重要作用,甲基结合蛋白(Methyl-Binding Domain Protein,MBD)是能特异识别甲基化位点的反式作用因子。开展植物MBD基因的功能研究对于探讨植物生长发育的表观遗传学调控机制具有重要意义。该研究构建了小麦甲基结合蛋白基因MBD3的原核表达载体pGEX-4T-MBD3,转化大肠杆菌BL21(DE3)工程菌株,在37℃,1mMIPTG浓度条件下,成功诱导表达了的GST-MBD3融合蛋白,大小为49.6kDа,这为进一步开展MBD3的蛋白纯化和功能分析奠定了基础。The DNA methylation play an important role in regulation of gene expression, methyl-binding domain protein is a kind of trans-acting factor which binds to methylated DNA. The functional analysis of MBD is a critical step toward understanding the epigenetic regulation mechanism on plant development. In this study, a prokaryotic expression vector pGEX-4T-MBD3 for the wheat MBD gene MBD3 was constructed. Transform it to E. eoli. BL21 （DE3）, the fusion protein GST-MBD3 with molecular weight of 49.6kDa was effectively expressed under lmM IPTG concentrations at 37℃, which provides a foundation for further purification and functional study of the MBD3 protein.

关 键 词：小麦MBD3 载体构建 原核表达 

分 类 号：Q784[生物学—分子生物学]