Cloning and Expressing Recombinant Protective Antigen Domains of B. anthracis

Report No. ARL-TR-5759
Authors: Deborah A. Sarkes, Joshua M. Kogot, Irene Val-Addo, Dimitra N. Stratis-Cullum, and Paul M. Pellegrino
Date/Pages: September 2011; 26 pages
Abstract: 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.
Distribution: Approved for public release
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Last Update / Reviewed: September 1, 2011