Molecular Cloning ofβ-Isopropylmalate Dehydrogenase Gene fromClostridium butyricumM588

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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 EcoRl 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 EcoRl fragments, was isolated from the transformants. It was found that the 2.2 kb EcoRl fragment on a reconstructed plasmid pCEl contained the β-isopropylmalate dehydrogenase gene. Restriction analysis showed that the β-isopropylmalate dehydrogenase gene was located in the 1.4kb Bglll-EcoRI fragment and the BamHl site was inside the β-isopropylmalate dehydrogenase gene.