Production of functional chick liver HMG 2a protein in Escherichia coli

FEBS Letters

Sasakawa, Takayo²  Kuwahata, Masashi²  Natori, Yasuo²  Horiuchi, Saburou¹  Oka, Tatsuzo²  Sassa, Toshihiro²  Miyamoto, Ken-ichi² 

[1] Department of Biochemistry, Iwate Medical University, Morioka, Iwate, Japan;Department of Nutrition, School of Medicine, The University of Tokushima, Tokushima, Japan

关键词: HMG 2a (chick liver);    Expression vector;    Recombinant HMG 2a;    Purification;    Escherichia coli;   

DOI  :  10.1016/0014-5793(95)00502-Z

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An efficient Escherichia coli system for the production of a variant form of high-mobility group-2a protein (HMG 2a), having the additional 5 amino acid residues (Ala-Pro-Thr-Leu-Glu) at the NH₂-terminal, has been constructed. cDNA encoding HMG 2a was ligated with the Omp A signal peptide sequence and was inserted into an inducible bacterial expression vector pSH-L. After the plasmid introduced into E. coli was expressed by temperature shift, the recombinant product was purified by trichloacetic acid precipitation followed by Bio-Rex 70 column chromatography. The purified product showed the expected NH₂-terminal sequence and the superhelical activity of circular DNA similar to the authentic HMG 2a isolated from chick liver.

【 授权许可】

Unknown