Cloning and Expressing Recombinant Protective Antigen Domains of B. anthracis
Rept. for Oct 2010-Jul 2011
ARMY RESEARCH LAB ADELPHI MD SENSORS AND ELECTRON DEVICES DIRECTORATE
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Protective antigen PA of Bacillus anthracis is a four-domain protein that is required for transport of edema factor EF and lethal factor LF into host cells during anthrax infection. Each of the four domains of PA plays a role in infection host cell receptor binding, domain 4, PA heptamerization and host membrane insertion, domains 2 and 3, and EF and LF recognition and intracellular transport, domain 1. This report details our efforts to produce single- and multi-domain combinations of the PA protein. All multi-domains 1-2, 2-3, 3-4 and three of the four single domains 1, 2, 4 have been cloned, expressed, and purified and will be used for future efforts in domain mapping of PA-binding antibodies and peptide reagents.
- Genetic Engineering and Molecular Biology