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 

出　　处：《Science China(Life Sciences)》1998年第5期548-554,共7页中国科学（生命科学英文版）

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

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.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.

关 键 词：TGFΒ1 E. COLI EXPRESSION in CYTOPLASM assembly and RENATURATION in vitro. 

分 类 号：Q987[生物学—遗传学]