Ligation, Transformation and Characterization of Rv 1926c Mycobacterium tuberculosis to Escherichia coli JM 109 For Latent Tuberculosis Immunodiagnostic

R. Agus
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Abstract

Tuberculosis caused by Mycobacterium tuberculosis is the biggest infectious disease causing human death in the world. The main challenge in controlling tuberculosis is to quickly and accurately diagnose tuberculosis infection. Several kits have been produced to diagnose tuberculosis, but have different sensitivity and specificity. This shows that the kit is not yet ideal for diagnosing tuberculosis, so the search for candidates for specific antigens still needs to be done. One potential antigen is the Rv 1926c encoding MPT 63 protein. This protein is known to induce Th1 cells and produce IFN λ from PBMC cells of patients infected with tuberculosis. The purpose of this study was to clone the Rv 1926c from Mycobacterium tuberculosis as a tuberculosis immunodiagnostic kit. The method used is isolating Rv 1926c with PCR, ligation to pGEM-T vector and transformation to E.coli host cell JM 109. Clone characterization was carried out by PCR and migration analysis. The results obtained are the recombinant clones obtained have successfully inserted with the Rv 1926 c
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结核分枝杆菌Rv 1926c与大肠杆菌JM109的连接、转化及特性研究
由结核分枝杆菌引起的结核病是世界上导致人类死亡的最大传染病。控制结核病的主要挑战是快速准确地诊断结核病感染。已经生产了几种诊断结核病的试剂盒,但灵敏度和特异性不同。这表明该试剂盒尚不适合诊断结核病,因此仍需寻找特异性抗原的候选者。一种潜在的抗原是编码MPT63蛋白的Rv 1926c。已知该蛋白可诱导Th1细胞,并从感染结核病的患者的PBMC细胞中产生IFNλ。本研究的目的是从结核分枝杆菌中克隆Rv 1926c作为结核病免疫诊断试剂盒。所使用的方法是用PCR分离Rv 1926c,连接到pGEM-T载体并转化到大肠杆菌宿主细胞JM 109。通过PCR和迁移分析进行克隆鉴定。所获得的结果是所获得的重组克隆已成功插入Rv 1926 c
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