Cloning the mutated immune antigen of Bacillus Anthracis in Bacillus subtilis

Aim And The immune antigen of Bacillus anthracis is a protein that can attach to the surface receptor of all human cells. At the surface of cancer cells there is a receptor that activates the uPA (Urokinase plasminogen) that do not exist in normal human cells. This research was conducted to change the site of attachment of PA gene with making direct mutation so that it can attach only to the cancer cells. In this study, pMNA1 plasmid containing PA gene was extracted from its host by basic lyse. Subsequently, phosphorylized primers and Overlap Extention PCR were used to make mutation on PA gene. The mutated segment was directly cloned in PTZ57R carrier and transferred to DH5α strain of E. coli. By catalyst enzymes KpnI and SalI the mutated PA gene was extracted and transferred to pWB980and by electroporation method it was transferred to WB600 strain. In this study mutation was occurred in sequences of PA gene by SOE PCR method resulting in a change in the genetic code of amino acid 194. The occurrence of mutation was confirmed by determining of base sequences. Cancer is a serious disease that in many cases leads to death. One of the treatment methods of cancer is bacterial toxins if only cancer cells receive them. Therefore, using changed PA can be a hope in cancer treatment as it attaches only to cancer cells.