HIGH LEVELS OF CONSTRUCTIVE PRODUCING ANTHRAX PROTECTIVE ANTIGEN

摘要

FIELD: biotechnology, microbiology, genetic engineering, molecular biology.;SUBSTANCE: invention relates to a method for preparing anthrax protective antigenic protein (PA). Protective antigenic protein is prepared by using the modified E. coli culture in its culturing with feeding. E. coli cells are transformed with the recombinant constitutive expression plasmid DH5 comprising PA gene to obtain E. coli DH5 cells expressing the protein PA. After culturing high density cells are selected, solubilized with 6-8 M urea and separated by centrifugation. Urea-denaturated PA is isolated from supernatant, purified and eluted. Also, method describes anthrax PA as 6X-histidine-fused protein prepared by this method. In culturing method involves using the primary additive chosen from the group consisting of polythiol, carbohydrate and organic acid, and LB medium (Luria broth) is used for feeding beginning from middle log-phase. As a plasmid for transformation of E. coli cells method involves vector of series pQE comprising phage promoter recognized by E. coli cells. Using the invention provides enhancing the level for producing anthrax protective antigen wherein antigen is obtained in form of protein fused with 6X-histidine.;EFFECT: improved preparing method, high level of antigen.;13 cl, 3 dwg, 1 tbl