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 journalArticle

3 Citations (Scopus)

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 overexposed 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
DNA-Binding Proteins
Bacilli
Escherichia coli
Organism Cloning
Column chromatography
Recombinant Proteins
Genes
Recombinant proteins
Agarose Chromatography
Proteins
Ion Exchange Chromatography
Circular Dichroism
Bacillus subtilis
Ribosomes
Sepharose
Open Reading Frames
Amino Acid Sequence

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 journalArticle

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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