Molecular cloning and analysis of the argC gene from Corynebacterium glutamicum

Abstract

The argC gene encoding N-acetylglutamate 5-semialdehyde dehydrogenase has been cloned from Corynebacterium glutamicum by transforming Escherichia coli arginine auxotroph with the genomic DNA library. Based on the restriction map of the cloned DNA, we have subcloned and sequenced the minimal DNA fragment complementing the E. coli argC mutant. The coding region of the cloned gene is 1041 nucleotides long with a predicted molecular mass of about 38 kDa polypeptide. Enzyme activity and size of the expressed protein in the E. coli auxotroph carrying the recombinant argC gene revealed that the cloned gene indeed codes for N-acetylglutamate 5-semialdehyde dehydrogenase. Computer analysis of the amino acid sequences of the predicted protein revealed a strong similarity to the corresponding protein of other bacteria.