HCV核心基因转染HepG2细胞后基因表达差异筛选.pdfVIP

● ● 2009 1 1 12 论著 ， 530028 ） 将真核表达载体pcDNA3.1 （－）HCV core转染到HepG2细胞，在HepG2细胞中表达HCV 核心蛋白，并筛选其中的差异表达基因。将构建的真核表达载体pCDNA3.1 （－）HCV core转染 HepG2细胞后进行蛋白免疫印迹检测；将pcDNA3.1 （－）HCV core和pcDNA3.1 （－）载体分别转染 HepG2细胞后，提取mRNA并逆转录为cDNA，运用基因表达谱芯片技术分析差异表达基因。构 建的真核表达载体经双酶切鉴定；转染HepG2细胞后HCV核心蛋白表达经蛋白免疫印迹证实；经基 因表达谱芯片分析发现，其中基因表达水平显著上调和下调的分别是181个和48个。筛选HCV核 心基因转染HepG2细胞后的糖类和脂类物质代谢相关的差异表达基因，从而为丙型肝炎病毒合并糖尿 病、脂肪肝等代谢性疾病的分子生物学机制的研究提供了重要依据。 丙型肝炎病毒核心抗原；HepG2细胞；基因芯片；差异表达基因 Screening of genes differentially expressed in HepG2 cells transfected with HCV core gene TANG Zhi-rong, CHEN Jie, LU Hong-yan, HU Yue-ying (Guangxi Center for Diseases Prevention and Control, Nanning 530028, China) Abstract: Objective To detect HCV core expression and screen different gene expression in HepG2 cell transfected by HCV core by genechip technology. Methods The expression vector of pcDNA3.1(-) HCV core is transfected with HepG2 cell line. The expression of HCV core protein was observed by Western blot method. Comparison of differentially expressed genes between L02 transfected by pcDNA3.1(-) HCV core and pcDNA3.1(-) by cDNA microarray technique was carried out, respectively. Results The expression vector has been confirmed by restriction enzyme digestion. HCV core protein expression has been confirmed by Western blot. High quality mRNA and cDNA had been prepared and successful microarray screening had been conducted. From the scanning results, it was found 181 genes were up-regulated and 48 genes were down-regulated in HepG2 cell line transfected with HCV core. Conclusions cDNA microarray technology was successfully applied to screen th