Cloning of the gene encoding dna binding protein hu from bacillus stearothermophilus and its expression in escherichia coli

Shunsuke Kawamura, Hirofumi Kajiyama, Nobuyuki Yamasaki, Makoto Kimura

Research output: Contribution to journalComment/debate

Abstract

The gene (hbst) encoding the DNA binding protein HU from Bacillus stearothermophilus was cloned, with the Bacillus subtilis HU gene (hbsu) as a hybridization probe. The nucleotide sequence, which contains a ribosome binding site, a transcriptional termination signal, as well as the coding region, was analyzed by the dideoxy chain-termination method. The deduced amino acid sequence of an open reading frame was perfectly matched with that of B. stearothermophilus HU (BstHU) determined by the protein chemical methods [M. Kimura and K. S. Wilson, J. Biol. Chem., 258, 4007-4011 (1983)]. The gene, hbst, was overexpressed using the expression vector pET-5a in Escherichia coli, and the recombinant HU protein (r-BstHU) was purified to be homogeneity by heparin-agarose column chromatography followed by ion-exchange column chromatography on S-Sepharose. The recombinant protein thus obtained had a circular dichroism spectrum identical to that of the authentic protein and bound to DNA to the same extent as the authentic protein.

Original languageEnglish
Pages (from-to)126-129
Number of pages4
JournalBioscience, Biotechnology and Biochemistry
Volume59
Issue number1
DOIs
Publication statusPublished - Jan 1 1995

Fingerprint

Geobacillus stearothermophilus
Gene encoding
Cloning
Bacilli
Escherichia coli
Organism Cloning
Carrier Proteins
Column chromatography
Recombinant Proteins
Genes
Recombinant proteins
Agarose Chromatography
Proteins
Ion Exchange Chromatography
DNA-Binding Proteins
Circular Dichroism
Bacillus subtilis
Ribosomes
Sepharose
Open Reading Frames

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Cloning of the gene encoding dna binding protein hu from bacillus stearothermophilus and its expression in escherichia coli. / Kawamura, Shunsuke; Kajiyama, Hirofumi; Yamasaki, Nobuyuki; Kimura, Makoto.

In: Bioscience, Biotechnology and Biochemistry, Vol. 59, No. 1, 01.01.1995, p. 126-129.

Research output: Contribution to journalComment/debate

Kawamura, Shunsuke ; Kajiyama, Hirofumi ; Yamasaki, Nobuyuki ; Kimura, Makoto. / Cloning of the gene encoding dna binding protein hu from bacillus stearothermophilus and its expression in escherichia coli. In: Bioscience, Biotechnology and Biochemistry. 1995 ; Vol. 59, No. 1. pp. 126-129.
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