Mukhija R, Rupa P, Pillai D, Garg L C
Gene Regulation Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India.
Gene. 1995 Nov 20;165(2):303-6. doi: 10.1016/0378-1119(95)00525-b.
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 His6 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 Ni(2+)-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 (Nb2) cell bioassay.
构建了一种质粒，用于在大肠杆菌中指导重组人生长激素（re-hGH）的合成，作为一种在T5启动子控制下N端含有His6标签的融合蛋白。大量合成了re-hGH，用异丙基-β-D-硫代半乳糖苷（IPTG）诱导后以包涵体形式积累。包涵体在6M盐酸胍中溶解，re-hGH通过在镍（2+）-次氮基三乙酸（NTA）琼脂糖上的单步亲和层析进行纯化。在摇瓶水平上，获得了纯化的re-hGH，产量为每升培养物30mg。re-hGH在大鼠淋巴瘤（Nb2）细胞生物测定中具有生物活性。
