HIGH-LEVEL PRODUCTION AND ONE-STEP PURIFICATION OF BIOLOGICALLY-ACTIVE HUMAN GROWTH-HORMONE IN ESCHERICHIA-COLI

被引：41

作者：

MUKHIJA, R

RUPA, P

PILLAI, D

GARG, LC

机构：

[1] NATL INST IMMUNOL,GENE REGULAT LAB,NEW DELHI 110067,INDIA

[2] UNIV DELHI,DEPT ZOOL,NEW DELHI,INDIA

来源：

GENE | 1995年 / 165卷 / 02期

关键词：

AFFINITY CHROMATOGRAPHY; BIOASSAY; CLONING; HISTIDINE TAG; INCLUSION BODY; RECOMBINANT DNA;

D O I：

10.1016/0378-1119(95)00525-B

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A plasmid has been constructed to direct the synthesis of recombinant human growth hormone (re-hGH) in Escherichia coli as a fusion protein containing a His(6) tag at the N-terminus under the control of the T5 promoter. The re-hGH was synthesized in large amounts and accumulated in the form of inclusion bodies upon induction with IPTG. Inclusion bodies were solubilized in 6 M guanidine-HCl and the re-hGH was purified by single-step affinity chromatography on Ni2+-nitrilotriacetic acid (NTA) agarose. At the shake flask level, the purified re-hGH was obtained with a yield of 30 mg/l of culture. The re-hGH was biologically active in a node rat lymphoma (Nb-2) cell bioassay.

引用

页码：303 / 306

页数：4

共 19 条

[1] EXPRESSION OF GROWTH HORMONE-RELEASING FACTOR ANALOG LEU27GRF(1-44)OH IN ESCHERICHIA-COLI - PURIFICATION AND CHARACTERIZATION OF THE EXPRESSED PROTEIN [J].