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
收藏
页码：303 / 306
页数：4
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