摘要
目的 建立肝脏持续表达丙型肝炎病毒核心 (HCVcore)蛋白的大鼠模型 ,为进一步研究HCV核心蛋白的功能和体内致病机制提供帮助。方法 应用聚合酶链反应法扩增出HCV Core区cDNA ,以重组腺伴随病毒载体介导的病毒重组技术制备含HCVCore的重组腺伴随病毒。重组病毒感染 2 0只雄性SD大鼠肝脏 ,感染后 1个月及 4个月行Southernblot印迹杂交法和Dotblot杂交法检测鼠肝内HCVCore区基因整合、mRNA形成情况。结果 测序证实成功克隆出HCVCorecDNA ,重组腺伴随病毒滴度为 2 .41× 10 11/ml。感染后 1个月及 4个月检测 ,实验大鼠肝脏Southernblot杂交结果均为阳性 ,HCVCore区基因非定点整合 ;肝细胞mRNA的Dotblot杂交结果均为阳性 ,提示正确表达HCVCoremRNA。结论 成功制备出一种肝脏持续表达HCVcore蛋白的大鼠模型。
Objective To establish a bandicoot model expressing hepatitis C virus (HCV) core protein persistently in liver for the help of farther study of the function and pathogenic mechanism of HCV core in vivo.Methods The cDNA coding for the HCV core was amplified by using polymerase chain reaction (PCR).The recombinant adenoassociated virus (rAAV) containing HCV core were prepared by using recombinant virus technology.Then the livers of 20 SD bandicoots were infected by rAAV.At the first and 4th month,each 10 bandicoots were anatomised for detection.Southern blot assay and Dot blot assay were used to detect the integrated HCV core genes and HCV core mRNA respectively.Results The cloned HCV core cDNA was confirmed successfully by using sequence analysis.The titer of the rAAV was 2.41×10 11 /ml.One and 4 months after infection,all the results of Southern blot assay and Dot blot assay were positive,indicating that the integrated HCV core genes and HCV core mRNA existed in the liver of bandicoots.Conclusion A bandicoot model expressing hepatitis C virus core protein persistently in liver was established successfully.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2003年第3期273-274,共2页
Chinese Journal of Experimental Surgery
