摘要
SARS冠状病毒核心蛋白对hfgl2纤维介素基因有激活作用,采用实时荧光定量PCR和Western印迹已验证了hfgl2基因在mRNA和蛋白水平的表达.为进一步明确在SARS冠状病毒核心蛋白刺激下,hfgl2纤维介素基因5′端非编码区对转录激活起重要作用的转录调控序列,构建了一系列hfgl2基因启动子荧光素酶报告基因质粒,将其与SARS冠状病毒核心蛋白真核表达质粒共转染CHO细胞.结果表明,转染前3个质粒hfgl2p(-1334)LUC、hfgl2p(-997)LUC、hfgl2p(-816)LUC的细胞的相对荧光素酶活性无显著改变;但转染hfgl2p(-468)LUC质粒的细胞荧光素酶的活性较前明显降低,说明在hfgl2基因启动子-816位至-468位(相对于转录起始点)之间存在着激活该基因的调控序列.本研究在一定程度上从分子水平揭示了SARS冠状病毒蛋白与宿主纤维介素基因之间的关系.
Using real-time fluorescence quantitative RT-PCR and Western blot, We demonstrate that the nucleocapsid（N） protein of SARS-CoV induces transcription of the novel fgl2 prothrombinase gene. A series of hfgl2 promoter/luciferase reporter constructs containing progressive deletions of hfgl2 promoter within the 5＇ flanking uncoding region were cotransfected with a nucleocapsid protein expressing construct in Chinese hamster ovary （CHO） ceils. There was no significant difference in luciferase activity in ceils cotransfected with hfgl2p （- 1334）LUC, hfgl2p （- 997）LUC and hfgl2p （- 816）LUC. However, Relative luciferase activity cotransfected with hfgl2p（ -418）LUC showed a remarkable decline compared with the other three constructs. Preliminary mapping of the hfgl2 promoter has defined a region from - 816 to - 468 （relative to the transcriptional start site） to be responsive for transcription regulation of hfgl2 gene in response to nucleocapsid protein. These results reveal the regulatory elements from - 816 to - 468 in hfgl2 promoter account for the activation of hfgl2 gene in response to nucleocapsid protein of SARS-CoV and show some light on the relation between SARS-CoV protein and host hfgl2 gene.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2007年第2期130-135,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
科技部973-SARS专项(No2003CB514100)基金
教育部首批SARS项目基金(教技司200364号)资助~~
