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

IF 3.5 Q3 Biochemistry, Genetics and Molecular Biology Journal of Genetic Engineering and Biotechnology Pub Date : 2018-12-01 DOI:10.1016/j.jgeb.2018.04.001
Ahyar Ahmad , Rosana Agus , Muh. Nasrum Massi , Rosdiana Natzir , Radha Madhyastha , Harish Kumar Madhyastha , Masugi Maruyama
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引用次数: 4

Abstract

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.

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结核分枝杆菌MPT83基因在大肠杆菌BL21中克隆及表达的初步研究
导致耐药性的结核分枝杆菌菌株的出现给世界各地的结核病治疗带来了新的问题,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,相信开发更有效的疫苗将阻止结核病的流行。一些结核分枝杆菌抗原,其中一种是MPT83,已经作为结核疫苗候选物进行了研究。MPT83抗原在脂蛋白微细菌中具有很强的免疫原性,经细胞计数循环鉴定为与抗原相关的表面细胞。MPT83对卡介苗具有结核病耐药性,被认为是可保护成年人群免受结核病侵害的结核病候选疫苗。本研究的目的是对MPT83抗原进行扩增克隆,并在大肠杆菌上表达其抗原。从研究结果来看,有望获得生产结核病疫苗的原料和高质量的抗原。PCR产物中的DNA条带为660 bp,而pGEMT-Easy-Mpt83重组质粒中的DNA条带为3678 bp。该蛋白在大肠杆菌BL21中表达,产生48 kDa蛋白和GST-MPT83融合蛋白。
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来源期刊
Journal of Genetic Engineering and Biotechnology
Journal of Genetic Engineering and Biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
5.70
自引率
5.70%
发文量
159
审稿时长
16 weeks
期刊介绍: Journal of genetic engineering and biotechnology is devoted to rapid publication of full-length research papers that leads to significant contribution in advancing knowledge in genetic engineering and biotechnology and provide novel perspectives in this research area. JGEB includes all major themes related to genetic engineering and recombinant DNA. The area of interest of JGEB includes but not restricted to: •Plant genetics •Animal genetics •Bacterial enzymes •Agricultural Biotechnology, •Biochemistry, •Biophysics, •Bioinformatics, •Environmental Biotechnology, •Industrial Biotechnology, •Microbial biotechnology, •Medical Biotechnology, •Bioenergy, Biosafety, •Biosecurity, •Bioethics, •GMOS, •Genomic, •Proteomic JGEB accepts
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