摘要
将中国人丙型肝炎病毒（ＨＣＶ）分离株的５’－非编码区和结构基因（５ＵＴＲ＋Ｃ＋Ｅ１＋Ｅ２）区基因插入到ｐｃＤＮＡ３载体中构成表达质粒，通过显微注射法将此质粒注射入Ｆ１代杂交鼠（Ｃ５７ＢＬ／６ＸＩＣＲ）受精卵，并植入假孕母鼠输卵管中，经筛选获得整合有目的基因的转基因小鼠系。免疫组化研究表明，ＨＣＶ核壳蛋白在转基因小鼠各组织中皆有表达，多呈核型染色，心肌细胞中可见浆型。核壳蛋白在各组织中的表达水平；心＞肺＞肾＞肝，同时在转基因小鼠心、肾、肝中检出Ｆａｓ抗原的表达，其表达水平与核壳蛋白的表达呈正相关关系。提示ＨＣＶ核壳蛋白的表达可引起Ｆａｓ抗原的表达。
Hepatitis C virus (HCV) is a leading cause of morbidity and mortality worldwide but the pathogenesis of liver cell injury and the molecular mechanism of persistent infection of HCV are not well understood. The aim of this work is to establish the animal model of HCV infection for studying the biosynthesis and pathogenesis of HCV structural protein in transgenic mice. The structural genes (C+E1+E2) and 5'-uncoding region gene (5'UCR) of HCV isolated from China was inserted into pcDNA3 vector. The transgenic mice were produced by microinfecting this plasmid into mouse (C57BL/6×ICR) embryos and the integrated mice were identified. The expression of core protein of HCV and mFas molecule were confirmed by immunohistochemical analysis using monoclonal antibody. The results revealed that the expression of core protein was detected in many mouses tissues with nuclear staining, while the heart tissue was with cytoplasmic staining. The expression level in tissues was indicated as: heart>lung>kidney>liver. At meantime, Fas molecule was detected in heart、kidney and liver tissues, and the expression of Fas was parallel to that of core protein. Transgenic mouse containing HCV structural gene was established and the expression of Fas molecule may be associated with the expression of core protein of HCV. The Fas system or other signaling pathway to the programmed cell death may play important roles in HCV infection. Our transgenic mouse will provide a useful animal model to critically address issues of immune pathogenesis、direct cytopathic potential and carcinogenesis of the structural protein of HCV.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
1997年第3期205-207,共3页
Chinese Journal of Experimental and Clinical Virology
关键词
丙型肝炎病毒
病毒蛋白质
转基因小鼠
丙型肝炎
Hepatitis C virus
Viral protein
△Fas molecule
△Transgenic mice
Gene
viral
