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Production of functional chick liver HMG 2a protein in Escherichia coli
Author(s) -
Oka Tatsuzo,
Sasakawa Takayo,
Miyamoto Ken-ichi,
Kuwahata Masashi,
Sassa Toshihiro,
Horiuchi Saburou,
Natori Yasuo
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00502-z
Subject(s) - escherichia coli , complementary dna , recombinant dna , signal peptide , peptide sequence , plasmid , biochemistry , biology , amino acid , expression vector , microbiology and biotechnology , chemistry , dna , gene
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 NH 2 ‐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 NH 2 ‐terminal sequence and the superhelical activity of circular DNA similar to the authentic HMG 2a isolated from chick liver.