Expression of human transforming growth factor β1 in E. coli (I) --Direct expression in cytoplasm

作     者：龙建银 王会信 刘莉 王芳 周廷冲 

作者机构：Beijing Institute of Basic Medical Sciences China 

出 版 物：《Science China(Life Sciences)》 (中国科学（生命科学英文版）)

年 卷 期：1998年第41卷第5期

页      面：548-554页

学科分类：0710[理学-生物学] 07[理学] 071007[理学-遗传学] 

基　　金：ProjectsupportedbytheNationalNaturalScienceFoundationofChina (GrantNo .395 80 0 15 ) 

主　　题：TGFβ1 E. coli expression in cytoplasm assembly and renaturation in vitro. 

摘      要：PCR technique is used to amplify the mature peptide gene of human transforming growth factor β1(hTGFβ1); the gene is verified by full length sequence analysis. In DH5α/pBV220 expression system, hTGFβ1 attains expression in the cytoplasm of \%E. coli\% up to 16%. The recombinant protein is proved to be the monomer of hTGFβ1 by N terminal amino acids analysis and immunoblotting. After refolding of the monomer protein \%in vitro\% in glutathione system or CHPAS/DMSO system, the dimeric protein accumulates to 30% in the refolding mixture. The recombinant protein is purified to homogeneity on silver staining, and is shown to have strong biological activity from MTT bioassay on Mv1Lu cells.