摘要
目的:检测HBV pre-X在真核表达载体pcDNA3.1-myc-his-HBV pre-X转染的HepG2细胞中的表达,并筛选其中的代谢相关差异表达基因.方法:将构建的真核表达载体pcDNA3.1-myc-his-HBV pre-X转染HepG2细胞后蛋白免疫印迹检测;将pcDNA3.1-myc-his-HBV pre-X和pcDNA3.1-myc-his载体分别转染HepG2细胞后,提取mRNA后逆转录为cDNA,运用基因表达谱芯片技术分析差异表达基因.结果:构建的真核表达载体经ApaⅠ、BstⅪ双酶切鉴定,转染HepG2细胞后HBV pre-X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200个和62个.结论:筛选HBV pre-X转染HepG2细胞后的代谢相关差异表达基因,从而为乙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.
AIM：To detect the expression of HBV pre-X and screen the different gene expression in HepG2 cells transfected with HBV pre-X using gene chip technology.METHODS：The expressive vector of pcDNA3.1-myc-his-HBV pre-X was transfected into HepG2 cell line.The expression of HBV pre-X protein was observed by western-blotting method.cDNA microarray technique was applied to de-tect the differentially expressed genes in HepG2 transfected either with pcDNA3.1-myc-his-HBV pre-X or with pcDNA3.1-myc-his.RESULTS：The expressive vector was confirmedby restriction enzyme digestion.The expression of HBV pre-X protein was confirmed by west-ern-blotting method.High quality mRNA and cDNA was prepared and successful microarray screening was conducted.Based on the scanning results,200 genes were found to be up-regulated and 62 genes down-regulated in HepG2 cell line transfected with HBV pre-X.CONCLUSION：cDNA microarray technology is successfully used to screen the genes differen-tially expressed in HepG2 cell line transfected with HBV pre-X,which may bring some new clues for studying the molecular biology mecha-nism of hepatitis B virus with metabolic diseases.
出处
《世界华人消化杂志》
CAS
北大核心
2008年第33期3724-3728,共5页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30600524~~
