Cloning and expression of the Thiobacillus ferrooxidans 3-isopropylmalate dehydrogenase gene in Escherichia coli

Kenji Inagaki, Hiroshi Kawaguchi, Yasuyuki Kuwata, Tsuyoshi Sugio, Hidehiko Tanaka, Tatsuo Tano

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The 3-isopropylmalate (3-IPM) dehydrogenase [EC 1.1.1.85] gene leuB of an acidophilic autotroph Thiobacillus ferrooxidans was cloned and expressed in Escherichia coli. Recombinant plasmids pTFL1 and pTFL2, carrying a 6.7-kb PstI fragment in the opposite orientation, conferred the same level of 3-IPM dehydrogenase activity on an E. coli leuB mutant. Restriction endonuclease mapping and deletion analysis indicated that 3-IPM dehydrogenase gene was on a 3.1-kb PstI-NruI fragment. The expression of the 3-IPM dehydrogenase gene of T. ferrooxidans was repressed by the addition of leucine in the culture medium of E. coli carrying the plasmid pTFL1. These results indicate that the 6.7-kb fragment contains the leuB structural gene and its regulatory region.

Original languageEnglish
Pages (from-to)71-74
Number of pages4
JournalJournal of Fermentation and Bioengineering
Volume70
Issue number2
DOIs
Publication statusPublished - 1990

Fingerprint

3-Isopropylmalate Dehydrogenase
Thiobacillus
Cloning
Escherichia coli
Organism Cloning
Genes
Plasmids
Restriction Mapping
Nucleic Acid Regulatory Sequences
DNA Restriction Enzymes
Leucine
Culture Media
Oxidoreductases

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Cloning and expression of the Thiobacillus ferrooxidans 3-isopropylmalate dehydrogenase gene in Escherichia coli. / Inagaki, Kenji; Kawaguchi, Hiroshi; Kuwata, Yasuyuki; Sugio, Tsuyoshi; Tanaka, Hidehiko; Tano, Tatsuo.

In: Journal of Fermentation and Bioengineering, Vol. 70, No. 2, 1990, p. 71-74.

Research output: Contribution to journalArticle

Inagaki, Kenji ; Kawaguchi, Hiroshi ; Kuwata, Yasuyuki ; Sugio, Tsuyoshi ; Tanaka, Hidehiko ; Tano, Tatsuo. / Cloning and expression of the Thiobacillus ferrooxidans 3-isopropylmalate dehydrogenase gene in Escherichia coli. In: Journal of Fermentation and Bioengineering. 1990 ; Vol. 70, No. 2. pp. 71-74.
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AU - Tanaka, Hidehiko

AU - Tano, Tatsuo

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