Cloning and sequence determination of the aspartase-encoding gene from Brevibacterium flavum MJ233

A 2.5-kb EcoRI fragment containing the aspartase-encoding gene (aspA) of Brevibacterium flavum MJ233 was cloned into plasmid pUC18 using Southern hybridization with the Escherichia coli aspA gene as a probe. The complete nucleotide (nt) sequence of the cloned DNA indicated that the deduced gene product of the Br. flavum aspA is composed of 526 amino acids (aa). Comparison of the aa sequence to the corresponding sequences from E. coli, Bacillus subtilis and Pseudomonas fluorescens revealed 63, 47 and 57% homology, respectively. The aspA product was determined to have a size of approx. 57 kDa by SDS-PAGE.