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Cloning and expression of MPT83 gene from Mycobacterium tuberculosis in E. coli BL21 as vaccine candidate of tuberculosis: A preliminary study

机译:结核分枝杆菌结核分枝杆菌结核分枝杆菌MPT83基因的克隆与表达初步研究

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摘要

The appearance of Mycobacterium tuberculosis strains leading to drug resistance has caused new problems in TB treatment in various parts of the world and forces WHO to declare TB as a global emergency. With the increase of TB drug resistance, it is convinced that a more effective vaccine development will stop the epidemic of TB. Some M. tuberculosis antigens, one of which is MPT83, have been examined as TB vaccine candidate. MPT83 antigen, which is very immunogenic in lipoprotein micro bacteria, is identified as surface cell interrelated to antigen with cytometry circulation. Having TB resistance from BCG vaccine, MPT83 is considered TB vaccine candidate that can protect people against TB at adult age. The purpose of this research is to conduct amplification of MPT83 antigen cloning, and expression of its antigen on E. coli bacteria. From the result of the research, it is expected that raw material to produce TB vaccine as well as a high-quality antigen can be obtained. The band of DNA in PCR product is 660 bp, while the one in pGEMT-Easy-Mpt83 recombinant plasmid is 3678 bp. This is expressed in E. coli BL21 strain and produces 48 kDa protein as well as GST-MPT83 fusion protein.
机译:导致耐药性的结核分枝杆菌菌株的出现在世界各地引起了结核病治疗方面的新​​问题,并迫使世卫组织宣布结核病为全球紧急情况。随着结核病耐药性的增加,人们相信,更有效的疫苗研发将阻止结核病的流行。一些结核分枝杆菌抗原(其中之一是MPT83)已作为结核病疫苗候选者进行了检查。 MPT83抗原在脂蛋白微细菌中具有很高的免疫原性,经细胞计数循环鉴定为与抗原相关的表面细胞。 MPT83具有抗BCG疫苗的结核病抗性,被认为是可以保护成人抗结核病的结核病候选疫苗。这项研究的目的是进行MPT83抗原克隆的扩增,并在大肠杆菌中表达其抗原。根据研究结果,可以期望获得用于生产TB疫苗的原料以及高质量的抗原。 PCR产物的DNA条带为660 bp,而pGEMT-Easy-Mpt83重组质粒的DNA条带为3678 bp。它在大肠杆菌BL21菌株中表达,并产生48 kkDa蛋白以及GST-MPT83融合蛋白。

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