丙型肝炎病毒核心基因置换对病毒复制和感染影响的初步研究

黎刚; 曹明媚; 王文博; 任浩; 戚中田

微生物与感染 ›› 2010, Vol. 5 ›› Issue (3) : 135-139.

PDF(16894 KB)
欢迎访问《微生物与感染》官方网站,今天是
PDF(16894 KB)
微生物与感染 ›› 2010, Vol. 5 ›› Issue (3) : 135-139.
论著

丙型肝炎病毒核心基因置换对病毒复制和感染影响的初步研究

  • 黎刚; 曹明媚; 王文博; 任浩; 戚中田
作者信息 +

Preliminary study on influnce of core gene substitution in replication and infection of hepatitis C virus

  • LI Gang; CAO Ming-Mei; WANG Wen-Bo; REN Hao; QI Zhong-Tian
Author information +
文章历史 +

摘要

本文旨在探讨丙型肝炎病毒(HCV)核心(core)蛋白在病毒复制和感染中的作用,采取基因置换的方法,用HCV 1b型J4株的核心基因平行置换2a 型J6JFH1株的核心区,构建了FL-J6JFH/J4core嵌合复制子。体外制备RNA转录体,以脂质体介导转染Huh7.5.1肝癌细胞。转染后第5天,荧光定量PCR(FQ-PCR)方法检测细胞内的HCV RNA水平。第8天,以丙型肝炎患者血清为一抗,免疫荧光染色法(IFA)检测转染细胞内HCV蛋白表达。同时收集转染后第8天的细胞上清液,感染naïve Huh7.5.1肝癌细胞,72小时后 IFA检测HCV蛋白表达。结果显示,FL-J6JFH/J4core 转染组第5天细胞内RNA水平与野生型FL-J6JFH1接近,无显著差异(n=4,P>0.05)。免疫荧光检测发现,FL-J6JFH/J4core 转染细胞后第8天及其上清液感染naïve Huh7.5.1细胞的HCV阳性细胞数都低于野生型。本研究结果初步表明,HCV各株间核心基因的替换会影响HCV的蛋白翻译和感染性病毒颗粒的产生与释放。

Abstract

In order to explore the role of core protein on the replication and infection of hepatitis C virus (HCV), intergenotypic chimeric FL-J6JFH/J4 core replicon was constructed in which the core gene of Fl-J6JFH strain (genotype 2a) was replaced by corresponding gene from HC-J4 strain (1b). In vitro RNA transcripts of FL-J6JFH and FL-J6JFH/J4 core were prepared and transfected into Huh7.5.1 hepatocytes respectively with liposomes. At day 5 post-transfection, total cellular RNA was isolated and determined by fluorescence quantitative polymerase chain reaction (FQ-PCR) method. Immunofluorescence staining analysis was performed to test the expression of HCV proteins in transfected cells at day 8 post-transfection. The naïve Huh7.5.1 cells were inoculated with the supernatants collected at day 8 post-transfection, and HCV proteins were detected by immunofluorescence at 72 h after inoculation. The results demonstrated that, the intracellular HCV RNA values of FL-J6JFH/J4 core was close to that of wild-type FL-J6JFH1 at day 5 post-transfection, and no significant differences were observed (n=4, P>0.05). Immunofluorecene analysis showed that, as compared with that in the wild type cells, the expression of HCV proteins in either cells transfected with FL-J6JFH/J4 core RNA at day 8 post-transfection or inoculation with supernatants collected at day 8 post-transfection was significantly lower. The results suggest that the core gene substitution among different HCV strains might influence the HCV protein expression and the production of infectious viral particles.

关键词

丙型肝炎病毒 / 核心基因 / 复制 / 感染

Key words

Hepatitis C virus / core gene / Replication / Infection

引用本文

导出引用
黎刚; 曹明媚; 王文博; 任浩; 戚中田. 丙型肝炎病毒核心基因置换对病毒复制和感染影响的初步研究[J]. 微生物与感染. 2010, 5(3): 135-139
LI Gang; CAO Ming-Mei; WANG Wen-Bo; REN Hao; QI Zhong-Tian. Preliminary study on influnce of core gene substitution in replication and infection of hepatitis C virus[J]. Journal of Microbes and Infections. 2010, 5(3): 135-139

PDF(16894 KB)

Accesses

Citation

Detail

段落导航
相关文章

/