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

Abstract

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.

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Document Details

Document Type
Technical Report
Publication Date
Feb 01, 1990
Accession Number
ADA227442

Entities

People

  • Connie S Schmaljohn
  • Joel M. Dalrymple
  • Lauren C. Iacono-connors

Organizations

  • United States Army Medical Research Institute of Infectious Diseases

Tags

DTIC Thesaurus Topics

  • Albumins
  • Animals
  • Baculoviridae
  • Biological Toxins
  • Cell Physiological Processes
  • Cells
  • Chemistry
  • Genetic Structures
  • Glutamic Acid
  • Immunization
  • Lagomorphs
  • Lepidoptera
  • Microbiology
  • Proteins
  • Rodents
  • Vaccines
  • Viruses

Fields of Study

  • Biology

Readers

  • Microbial Pathology
  • Molecular and Cellular Biochemistry
  • Virology (or Medical Virology).