结核分枝杆菌MPT83基因在BL21中的克隆表达及作为结核病疫苗候选物的初步研究

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

作者信息

Ahmad Ahyar, Agus Rosana, Massi Muh Nasrum, Natzir Rosdiana, Madhyastha Radha, Madhyastha Harish Kumar, Maruyama Masugi

机构信息

Chemistry Department, Mathematic and Natural Science Faculty, Hasanuddin Univesity, Perintis Kemerdekaan Street km. 10 Tamalanrea, Makassar 90245, Indonesia.

Laboratory of Research Centre and Developing of Sciences, Faculty of Mathematic and Natural Sciences, Hasanuddin University, Jl. Perintis Kemerdekaan KM 10, Makassar 90245, Indonesia.

出版信息

J Genet Eng Biotechnol. 2018 Dec;16(2):335-340. doi: 10.1016/j.jgeb.2018.04.001. Epub 2018 Apr 13.

DOI:10.1016/j.jgeb.2018.04.001

PMID:30733743

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6353755/

Abstract

The appearance of 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 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 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 BL21 strain and produces 48 kDa protein as well as GST-MPT83 fusion protein.

摘要

导致耐药性的菌株出现给世界各地的结核病治疗带来了新问题，并迫使世界卫生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，人们确信开发更有效的疫苗将阻止结核病的流行。一些抗原，其中之一是MPT83，已被作为结核病疫苗候选物进行研究。MPT83抗原在脂蛋白微细菌中具有很强的免疫原性，通过细胞计数循环被鉴定为与抗原相关的表面细胞。由于卡介苗具有结核病抗性，MPT83被认为是一种可以在成人阶段保护人们免受结核病侵害的结核病疫苗候选物。本研究的目的是进行MPT83抗原克隆的扩增，并在细菌上表达其抗原。从研究结果来看，有望获得生产结核病疫苗的原料以及高质量的抗原。PCR产物中的DNA条带为660 bp，而pGEMT-Easy-Mpt83重组质粒中的条带为3678 bp。这在BL21菌株中表达，并产生48 kDa的蛋白质以及GST-MPT83融合蛋白。