نوع مقاله : مقاله پژوهشی
عنوان مقاله English
نویسندگان English
The basis for diagnosing cattle tuberculosis is the skin test or tuberculin test. However, the presence of false positive cases in this skin test due to the occurrence of cross-reactions of PPD prepared from the AN5 strain of Mycobacterium bovis with other mycobacteria is considered one of its disadvantages, which is why the use of proteins with higher specificity is recommended to overcome it. One of the objectives of this study was to clone, express, and purify the recombinant ESAT-6 protein and evaluate its immunogenicity as an important antigen of Mycobacterium bovis in the prokaryotic system. In this study, the esat-6 gene encoding the secretory protein ESAT-6 was amplified using the polymerase chain reaction method. Then, the obtained PCR product was digested with two restriction endonuclease enzymes EcoRI and HindIII and then cloned in the plasmid vector pET23a(+) and amplified after transformation in Escherichia coli DH5α. In order to express the recombinant ESAT-6 protein, the above vector containing the cloned esat-6 gene was transferred to Escherichia coli expressing BL21. The correctness of the gene cloning was confirmed by sequencing. In the following study, the recombinant ESAT-6 protein was purified using nickel resin and its antigenic expression was confirmed by Western blotting using specific monoclonal antibodies. The results of gene sequencing along with Western blotting indicated that the obtained recombinant protein was well expressed and purified and could be used in challenge experiments and diagnostic tests like the recombinant CFP-10 protein.
کلیدواژهها English
