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丙型肝炎病毒(HCV)感染患者HCV核心片段和NS3片段的分子克隆、测序及表达

Molecular cloning, sequencing and expression of core and NS3 fragments of HCV from patients with HCV infection.

作者信息

Tan D M, Hu G L, Arima T, Zhang Z, Nanba T, Hatanaka T

机构信息

Department of Infectious Diseases, First Affiliated Hospital, Hunan Medical University, Changsha.

出版信息

Chin Med J (Engl). 1993 Jul;106(7):522-6.

PMID:8243124
Abstract

Fragments of core and NS3 of hepatitis C virus-Hunan (HCV-Hun) were cloned by RT-PCR and gene recombinant techniques from blood samples collected in Hunan Province, China. In comparison with sequences of our samples with those of HCV-US and HCV-J, the homologies of nucleotides and amino acids were about 90%, indicating that fragments of core and NS3 of HCV-Hun were in a relative conserved region of HCV. Two fusion proteins containing the peptides coded by HCV core (MBP-HCV core) and HCV. NS3 (MBP-HCV. NS3-Gal) were expressed by Escherichia Coli with recombinant plasmids. The specific HCV antigenicity of the two fusion proteins were identified by western blotting. Therefore, MBP-HCV. core and MBP-HCV.NS3-Gal were found useful for anti-HCV assay.

摘要

通过逆转录聚合酶链反应(RT-PCR)和基因重组技术,从中国湖南省采集的血液样本中克隆出丙型肝炎病毒-湖南株(HCV-Hun)的核心片段和NS3片段。将我们样本的序列与HCV-US和HCV-J的序列进行比较,核苷酸和氨基酸的同源性约为90%,表明HCV-Hun的核心片段和NS3片段处于HCV的相对保守区域。用重组质粒通过大肠杆菌表达了两种包含由HCV核心编码的肽(MBP-HCV核心)和HCV NS3(MBP-HCV NS3-Gal)的融合蛋白。通过蛋白质印迹法鉴定了这两种融合蛋白的特异性HCV抗原性。因此,发现MBP-HCV核心和MBP-HCV NS3-Gal可用于抗HCV检测。

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