Expression of the 'Bacillus anthracis' Protective Antigen Gene by Baculovirus and Vaccinia Virus Recombinants
ARMY MEDICAL RESEARCH INST OF INFECTIOUS DISEASES FORT DETRICK MD
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The gene encoding Bacillus anthracis protective antigen PA was modified by site-directed mutagenesis, subcloned into baculovirus and vaccinia virus plasmid transfer vectors pAcYM1 and pSC-11, respectively, and inserted via homologous recombinations into baculovirus Autographa californica nuclear polyhedrosis virus or vaccinia virus strains WR and Connaught. Expression of PA was detected in both systems by immunofluorescence assays with antisera from rabbits immunized with B. anthracis PA. Western blot immunoblot analysis showed that the expressed product of both systems was slightly larger 86 kilodaltons than B. anthracis-produced PA 83.5 kilodaltons. Analysis of trypsin digests of virus-expressed and authentic PA suggested that the size difference was due to the presence of a signal sequence remaining with the virus-expressed protein.
- Medicine and Medical Research