S100A4原核表达载体的构建以及蛋白的表达和纯化  

Prokaryotic Expression and Purification of Bioactive Human S100A4

作　　者：韩雷[1] 何虹霖[1] 关文[1] 李京敬[1] 袁运生[1] 俞雁[1] 

机构地区：[1]上海交通大学农业与生物学院,上海200240

出　　处：《基因组学与应用生物学》2013年第2期142-148,共7页Genomics and Applied Biology

基　　金：上海科委国际合作项目支持

摘　　要：S100A4是S100蛋白家族的成员,在细胞的增殖、分化、损伤修复以及肿瘤细胞转移等方面发挥重要的调控作用。本研究将S100A4全长基因构建到pET28a原核表达载体上,利用大肠杆菌表达系统表达和纯化出高纯度的重组人S100A4。通过试验证明,重组人S100A4蛋白在体外可以有效地增强黑色素瘤细胞A375-S2的增殖。重组人S100A4原核表达与纯化方法的建立将促进其结构和生物学功能研究,并且对于S100蛋白家族其它蛋白的表达与纯化具有重要的参考意义。S100A4 is a member of S100 protein family, which exerts functions in cell proliferation, differention and cell damage repair. It is also reported that S 100A4 is assoicated with cancer metastasis. To facilitate further studies of S100A4, we developed a strategy for expressing and purifying recombinant human S100A4 protein using E. coli expression system and affinity chromotography methods. This strategy is simple and efficient for purifying recombinant S100A4 with high purity and low endotoxin contamination. Furthermore, in vitro studies validated the biological activity of the purified S 100A4 since it could enhance the viability of A375 cells. We hope that this strategy could facilitate the physiological and pathological research of S 100A4 and have reference values for the expression and purification of other S 100 family members.

关 键 词：S100A4 原核表达 蛋白纯化 蛋白活性 

分 类 号：Q78[生物学—分子生物学]