Cloning of the argF gene encoding the ornithine carbamoyltransferase from Corynebacterium glutamicum

Abstract

The argF gene encoding ornithine carbamoyltransferase (OTCase; EC2.1.3.3) has been cloned from Corynebacterium glutamicum by transforming the Escherichia coli arginine auxotroph with the genomic DNA library. The cloned DNA also complements the E. coli argG mutant, suggesting a clustered organization of the genes in the genome, me have determined the DNA sequence of the minimal fragment complementing the E, coli argF mutant. The coding region of the cloned gene is 957 nucleotides long with a deduced molecular mass of about 35 kDa polypeptide. The enzyme activity and size of the expressed protein in the E, coli auxotroph carrying the argF gene revealed that the cloned gene indeed codes for OTCase, Analysis of the amino acid sequence of the predicted protein revealed a strong similarity to the corresponding protein of other bacteria.