Mutation, cloning and sequencing of Protective Antigen of Bacillus anthrasis
Subject Areas : microbiologyمحمد ابوطالب 1 , Hatef Ajoudanifar 2
1 - دانشجوی دانشگاه آزاد اسلامی واحد قم
2 - عضو هیئت علمی دانشگاه آزاد اسلامی واحد دامغان
Keywords: Mutagenesis, Bacillus anthrasis, Protective Antigene,
Abstract :
Background: Protective Antigen of Bacillus anthracis (PA) is a protein that binds to receptors on OF human body cells. There is special Urokinase plasminogen activator receptor on cancer cells and PA can not bind to them. The aim of this study is modify the receptor of PA with site directed mutagenesis that the protein can only be bind to the cancer cells. Material and methods: In this study, pMNA1 plasmid that contained PA gene was extracted and site directed mutagenesis done with SOE PCR on PA gene. Mutant gene cloned on pTZ57R vector directly and transformed into E. coli DH5α with CaCl2 method. Finally exiting of the gene and mutation on PA evaluated with PCR , digestion and sequencing.Results: PA gene separated with PCR and mutated with SOE PCR. Mutated PCR product cloned on pTZ57R vector and earned 5.1 kb plasmid. Recombinant plasmid evaluated with digestion and PCR. Mutation confirmed with sequencing. Conclsion: Cancer has a deadly disease. One of the methods for treaeting cancer using bacterial toxins. Therefore Using a modified PA protein that binds to the cancer cells can create new hope for cancer treatment.
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