Expression of the 'Bacillus anthracis' Protective Antigen Gene by Baculovirus and Vaccinia Virus Recombinants

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 nuclea...

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Bibliographic Details
Main Authors: Iacono-Connors, Lauren C, Schmaljohn, Connie S, Dalrymple, Joel M
Format: Report
Language:English
Subjects:
ANTIBODIES
IMMUNE SERUMS
Medicine and Medical Research
RABBITS
SEQUENCES
SIGNALS
VACCINIA VIRUS
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Summary: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. Pub. in Infection and Immunity, v58 n2 p366-372 Feb 1990.