همسانه‌سازی و بیان ژن‌های ممزوجی دومن - a1 آنتی‌ژن حفاظتی با سیلوس آنتراسیس و زیرواحد B سم شیگلا در باکتری E. coli

Introduction: Shigella enterotoxin(STxB) is one of the major virulent factors in Shigella dysenteriae type 1 and E. coli O157:H7, which its immunogenicity, adjuvant and delivery characteristic have been proven. Anthrax is a common disease in humans and animals and identifies domain a-1 antibodies of protective antigen(PA) which is 62% of the PA antigens of Bacillus anthracis. Therefore, the purpose of this study was to investigate the expression of recombinant protein domain a-1 protective antigen(PA20) of Bacillus anthracis with Shigella enterotoxin B subunit(STxB) in E. coli. Methods: In this experimental study, primers of pa 20 gene were designed for replacement in stxB-ipaD gene cassette. PCR was performed in order to amplify the fragment and the amplified fragments were cloned into pGEM-Teasy vector. NdeI and SalI restriction enzymes cut pa20 genes and finally the genes pa20 were fused with genes stxB. PET28a (+) Vector containing the gene cassette pa20-stxB was transformed into E. coli strain BL21(DE3) and expression of gene cassette was studied. Results: pa20-stxB fused genes were confirmed in the expression vector pET28a(+) by PCR, enzyme digestion and Sequencing. The produced recombinant proteins were confirmed by SDS-PAGE and Western blotting. Conclusion: The findings of the current study revealed that this antigen can be raised as an anti-cancer and recombinant vaccine candidate against types of Shigella, Escherichia coli and Bacillus anthracis which can be due to such factors as identification of antigen(PA) by antibody PA20, its apoptosis induction properties, property of immunogenicity, adjuvant and delivery of STxB protein and high expression levels of Gb3 in human cancer cells.