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Asporogenic recombinant producer of anthrax protective antigen

Mikshis, N. I., Kudryavtseva, O. M., Shulepov, D. V., Goncharova, A. Yu., Bolotnikova, M. F., Novikova, L. V., Popov, Yu. A., Kutyrev, V. V.
Applied biochemistry and microbiology 2011 v.47 no.7 pp. 667-673
Bacillus anthracis, anthrax, antigens, hybrids, immunization, lethal dose 50, pathogenicity, plasmids, rabbits, structural genes
An asporogenic recombinant strain Bacillus anthracis 55ΔTPA-1(Spo−) producing anthrax protective antigen (PA) was obtained. The strain contains structural gene pag as a part of a hybrid replicon pUB110PA-1 and lacks determinants encoding the synthesis of main factors of anthrax pathogenicity. The level of PA production by asporogenic genetically engineered strain is approximately 80 μg/ml that is 4–5 times more than the values determined for vaccine strains B. anthracis STI-1 and B. anthracis 55. The strain preserves asporogenicity and ability to replicate the hybrid plasmid after in vitro passages. Biologically active PA was isolated from the constructed strain B. anthracis 55ΔTPA-1(Spo−). Double immunization of rabbits with 50 μg of the purified recombinant product provides their 100% protection from infection with 50 LD50 of a highly virulent anthrax strain.