人CXCL4蛋白原核表达与纯化  

Prokaryotic Expression and Purification of Bioactive Human CXCL4

作　　者：周亮[1] 叶浩[2] 周瓅 关文[1] 李京敬[2] 郜尽[2] 韩伟[2] 俞雁[1] 

机构地区：[1]上海交通大学农业与生物学院上海市兽医生物技术重点实验室,上海200240 [2]上海交通大学药学院再生组学课题组,上海200240

出　　处：《中国生物工程杂志》2016年第1期7-13,共7页China Biotechnology

基　　金：国家自然科学基金(81273573);上海市科学基金(11431921300)资助项目

摘　　要：CXCL4又名血小板因子4(PF4),属于CXC趋化因子亚家族,能够广谱的作用于多种细胞,在炎症,凝血等众多生理及病理反应中发挥重要作用。将CXCL4全长基因构建到原核表达载体p ET43.1a(+),利用大肠杆菌系统表达,继而纯化获得高纯度的目的蛋白。实验表明重组人CXCL4蛋白可以有效抑制人肾癌细胞增殖,具有生物学活性。重组人CXCL4原核表达与纯化方法的建立将有助于其生物学结构与功能的研究,并且对趋化因子家族其它蛋白质的表达与纯化具有参考价值。CXCL4,Platelet factor 4（ PF4）,belongs to the subfamily of CXC chemokines,which is reported to be involved in numerous biological process by acting on a broad spectrum of different cell types.Here,the study aimed to express and purify recombinant human CXCL4 protein using E.coli expression system and affinity chromotography methods.The results demonstrated a simple and efficient strategy for the production and purification of recombinant human CXCL4（ rh CXCL4） with over 95% purity by gel electrophoresis have been developed.Its biological activity was confirmed in the proliferation-inhibition assay using a tumor cell line,ACHN cells.The rh CXCL4 will facilitate the structural and functional analysis of human CXCL4.The production strategy of rh CXCL4 should have important reference values for the expression and purification of other chemokines.

关 键 词：CXCL4 原核表达 纯化 蛋白质活性 

分 类 号：Q493.2[生物学—生理学]