Molecular cloning of .BETA.-isopropylmalate dehydrogenase gene from Clostridium butyricum M588.

1 January 1983

journal article
Published by Oxford University Press (OUP) in Agricultural and Biological Chemistry

Vol. 47 (10) , 2313-2317
https://doi.org/10.1271/bbb1961.47.2313

Abstract

The gene for β-isopropylmalate dehydrogenase of Clostridium butyricum M588 has been cloned in E. coli. The genome of Clostridium butyricum M588 was digested with restriction endonuclease EcoRI and joined to plasmid pBR322. Competent E. coli HB101 cells were transformed with the hybrid plasmids and the leucine⁺ transformants were selected. The plasmid, pCE4, containing four EcoRI fragments, was isolated from the transformants. It was found that the 2.2kb EcoRI fragment on a reconstructed plasmid pCE1 contained the β-isopropylmalate dehydrogenase gene. Restriction analysis showed that the β-isopropylmalate dehydrogenase gene was located in the 1.4kb Bglll-EcoRl fragment and the BamRl site was inside the β-isopropylmalate dehydrogenase gene.

Keywords

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