Expression of Bacillus Anthracis Protective Antigen in Bacillus Megaterium

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Authors
  1. Berger, B.J.
  2. Schwandt, K.E.
  3. Radford, C.L.
Corporate Authors
Defence R&D Canada - Suffield, Ralston ALTA (CAN)
Abstract
Bacillus anthracis protective antigen is the central component of the anthrax toxin complexes that facilitates entry of lethal factor and edema factor into host cells. Protective antigen is also the major immunogenic component present in the currently licensed anthrax vaccine. In order to produce full-length, soluble protective antigen, the gene has been cloned and expressed using Bacillus megaterium and a xylose-inducible heterologous expression system. After only 3.5 hours growth post-induction in Luria-Bertani broth, the transgenic B. megaterium were found to secrete approximately 1 mu g/ml protective antigen into the culture medium. The recombinant protein was easily purified to homogeneity in a single step by ion exchange chromatography. N-terminal amino acid sequencing of the final product confirmed that the recombinant protective antigen was full-length and that no proteolytic degradation had occurred

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Keywords
Bacillus anthracis;Bacillus megaterium;Protective antigen;Recombinant protein;Purification;Amino acid sequencing
Report Number
DRDC-SUFFIELD-TM-2004-045 — Technical Memorandum
Date of publication
01 Mar 2004
Number of Pages
25
DSTKIM No
CA023870
CANDIS No
521389
Format(s):
Hardcopy;CD ROM

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