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缩短的人巨噬细胞集落刺激因子(M-CSF)在大肠杆菌中的表达
引用本文:董晨,华子春,陈于红,曾凡杰,朱洁. 缩短的人巨噬细胞集落刺激因子(M-CSF)在大肠杆菌中的表达[J]. Acta biochimica et biophysica Sinica, 1995, 0(3)
作者姓名:董晨  华子春  陈于红  曾凡杰  朱洁
作者单位:南京大学生物化学系,南京大学医药生物技术国家重点实验室
摘    要:本文用聚合酶链反应(PCR)获得了一个缩短的人巨噬细胞集落刺激因子(编码3~149氨基酸)cDNA基因,并克隆在质粒pET3d中,在T7启动子指导下,在大肠杆菌BL21(DE3)LysE中获得了和一个6组氨酸短肽标签的融合表达。重组的融合m-CSF表达量占菌体总蛋白的12%,表达产物一部分以不溶性包涵体形式存在,另一部分则以可溶性蛋白存在。经过金属螫合亲和层析一步纯化,所得的融合(His)6-M-CSF在还原型SDS-PAGE上基本呈一条均一的蛋白质条带。

关 键 词:人巨噬细胞集落刺激因子,金属螯合亲和层析,组氨酸短肽标签

^Expression of a Truncated Human Macrophage Colony-stimulating Factor in E. coli
DONG Chen, HUA Zi-Chun, OHEN Yu-Hong, ZENG Fan-Jie and ZHU Jie. ^Expression of a Truncated Human Macrophage Colony-stimulating Factor in E. coli[J]. Acta biochimica et biophysica Sinica, 1995, 0(3)
Authors:DONG Chen   HUA Zi-Chun   OHEN Yu-Hong   ZENG Fan-Jie  ZHU Jie
Abstract:A truncated cDNA of the human macrophage colony-stimulating factor gene, encoding amino acid residues from 3 to 149 of the native M-CSF, was isolated by using the polymerase chain reaction and inserted into plasmid pET3d. It directs the expression of truncated M-CSF fused with a six-histidine tail peptide in E. coli BL21(DE3) LysE under the control of T7 promoter. The expression level of the truncated M-CSF is twelve percent of total cellular proteins, with half of the product accumulated as inclusion bodies and the other part as soluble protein. Through the metalaffinity chromatography, the recombinant truncated M-CSF showed an almost homogenous protein band on reducing SDS-PAGE.
Keywords:Human macrophage colony-stimulating factor  Metalchelating affinity chromatography  Six-histidine tail peptide  
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