Use of Recombinant DNA Techniques for the Production of a More Effective Anthrax Vaccine
Abstract
We have continued to characterize the edema factor (EF) and lethal factor (LF) genes of Bacillus anthracis. Using site-specific mutagenesis, we have identified the ATP binding site of EF, which is a calmodulin-dependent adenylate cyclase. We have also identified a potential calmodulin binding site. In addition, we have shown that carboxyl terminus of EF shares at least three highly conserved amino acid domains with the calmodulin-dependent adenylate cyclase of Bordetella pertussis. We have also determined the DNA sequence of the LF gene. This gene contains many features in common with the EF and PA genes, including a strong ribosome binding site and a long signal peptide (33 amino acids) which is apparently removed during secretion. In addition, the amino terminal 300 amino acids of LF shares extensive homology with the amino terminus of EF. This shared homology is probably part of their binding domains for associating with PA since EF and LF each bind to PA. Each of the anthrax toxin genes (cya (EF), pag (PA) and lef (LF) have been expressed in Bacillus subtilis. In addition, expression plasmids have been constructed for regulated high level expression of these genes. We have also fused the EF coding region to the PA promotor and then expressed EF in B. subtilis using this plasmid construction. This plasmid has also been transferred into B. anthracis for high level, regulated expression. Expression of the individual toxin genes in B. subtilis should provide a safe bacterial host for production of large quantities of the B. anthracis toxin proteins.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 31, 1988
- Accession Number
- ADA201939
Entities
People
- Donald L. Robertson
Organizations
- Brigham Young University