摘要
目的:利用基因表达谱芯片技术筛选、比较丙型肝炎病毒NS5A反式调节基因2不同剪切体对HepG2细胞基因表达的调控作用及差异性.方法:利用分子生物学技术和生物信息学技术相结合,克隆人NS5ATP2的不同剪切体蛋白的编码基因,常规分子生物学技术构建相应的真核细胞表达载体pcDNA3.1-NS5ATP2(615)和pcDNA3.1-NS5ATP2(216).脂质体技术转染肝母细胞瘤细胞系HepG2利用基因表达谱芯片技术筛选转染细胞的差异表达cDNA的类型.结果:对于2个基因剪切体转染细胞系差异表达基因谱的分析,NS5ATP2(615)和NSSATP2(216)共同上调的基因43条;NS5ATP2(615)和NSSATP2(216)共同下调的基因13条;NS5ATP2(615)上调,而NS5ATP2(216)下调的基因类型3条;未发现NS5ATP2(615)下调;NS5ATP2(216)上调的基因类型;还有一些靶基因,或只受到NS5ATP2(615)的调节,或只受到NS5ATP2(216)的调节.结论:不同剪切体的作用之间有共同的靶基因,也有不同的靶基因,甚至对于同一基因有相反的调节功能.
AIM: To identify and analyze target genes regulated by different spliced transcripts of human NS5ATP2 by microarray technique. METHODS: The coding sequences for the two differently spliced transcripts of human NS5ATP2, NS5ATP2 (615) and NS5ATP2, were cloned into eukaryotic expression vector pcDNA3.1 to yield pcDNA3.1-NS5ATP2 (615)and pcDNA3.1-NS5ATP2 (216) constructs, which were then transfected into HepG2 cells. The differentially expressed genes in the cells transfected with the two differently spliced variants were analyzed and compared by DNA microarray. RESULTS: Forty-three genes were up-regulated by both NS5ATP2 (615) and NS5ATP2 (216), and 13 genes were down-regulated by both NS5ATP2 (615) and NS5ATP2 (216). Meanwhile, 3 genes were found to be up-regulated by NS5ATP2 (615), but down-regulated by NS5ATP2 (216). No genes were down-regulated by NS5ATP2(615) while up-regulated by NS5ATP2 (216).Many genes were trans-regulated by NS5ATP2 (615) or NS5ATP2 (216) independently. CONCLUSION: Different spliced transcripts of human NS5ATP2 may commonly or independently regulate target genes. They even exhibit opposite regulatory effects upon some target genes.
出处
《世界华人消化杂志》
CAS
北大核心
2005年第2期198-201,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金攻关项目
No.C03011402
No.C30070689
No C39970674
No.C30070689
No.C30371288军队九
五科技攻关项目
NO.98D063军队回国留学人员启动基金项目
No 98H038军队十
五科技攻关青年基金项目
No.01Q138军队十
五科技攻关面上项目
NO.01MBl35~~
