Javed F, Manzoor S, Khattak A Ali, Imran M, Parvaiz F, Ashraf J, Tariq H, Bhatti S
Acta Virol. 2015 Sep;59(3):284-94. doi: 10.4149/av_2015_03_284.
Hepatitis C virus (HCV) chronically infects almost 2% of world's population. Chronic infection can lead to liver failure and hepatocellular carcinoma (HCC). Approximately 10% of the Pakistani population is infected with HCV and type 3 is the most prevalent genotype with 75-90% prevalence. In this study we have developed transiently expressing cell culture based system for the expression of HCV non-structural NS3, NS3-4A and NS4A proteins of genotype 3a. HCV non-structural genes NS3, NS3-4A and NS4A were cloned in to pFLAG-CMV2 and pEGFP-C1vectors. All vectors were transfected separately to Huh-7 cells and their protein expression was analyzed by Western blot and immunofluorescence. All proteins were expressed correctly and in the transfection we have obtained 42-70% efficiency for all clones. This system can be used for the development of novel antiviral strategies to inhibit the viral replication, to study apoptosis pathways induced by HCV, for the evaluation of vaccine candidates and also to study the role of HCV different signaling pathways.
丙型肝炎病毒（HCV）慢性感染着全球近2%的人口。慢性感染可导致肝衰竭和肝细胞癌（HCC）。巴基斯坦约10%的人口感染了HCV，其中3型是最常见的基因型，患病率为75 - 90%。在本研究中，我们开发了基于瞬时表达细胞培养的系统，用于表达3a基因型HCV的非结构NS3、NS3 - 4A和NS4A蛋白。将HCV非结构基因NS3、NS3 - 4A和NS4A克隆到pFLAG - CMV2和pEGFP - C1载体中。将所有载体分别转染至Huh - 7细胞，并通过蛋白质印迹法和免疫荧光分析其蛋白表达。所有蛋白均正确表达，在转染中，所有克隆的效率为42 - 70%。该系统可用于开发新型抗病毒策略，以抑制病毒复制、研究HCV诱导的凋亡途径、评估候选疫苗，还可用于研究HCV不同信号通路的作用。
