Oka T, Sasakawa T, Miyamoto K, Kuwahata M, Sassa T, Horiuchi S, Natori Y
Department of Nutrition, School of Medicine, University of Tokushima, Japan.
FEBS Lett. 1995 Jun 19;367(1):49-52. doi: 10.1016/0014-5793(95)00502-z.
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.
