Cloning, sequence and expression of the gene (aprV5) encoding extracellular serine acidic protease V5 from Dichelobacter nodosus

Abstract

The acidic protease V5-encoding gene (aprV5) from Gram- Dichelobacter nodosus virulent strain 198 was isolated from a cosmid bank by activity screening and sequenced. The 2371-bp nucleotide (nt) sequence contained an open reading frame coding for a protein precursor of 595 amino acid (aa) residues composed of a signal peptide, a pro-region, a mature active protease of 347 aa and a C-terminal extension region of 120 aa. The deduced aa sequence of the pre-pro-mature protease regions showed about 65% similarity to that of D. nodosus basic protease while the C-terminal extension region showed only about 26% similarity. The aprV5 gene, without its C-terminal extension region, was cloned and expressed in Escherichia coli. The acidic protease B5-encoding gene (aprB5) from non-virulent strain 305 was also cloned and sequenced. The aprB5 nt sequence showed 99% homology to that of aprV5 with two single-aa changes occurring in the precursor.