SARS冠状病毒N蛋白激活hfgl2凝血酶原酶基因的研究

发布时间：2018-03-10 11:20

本文选题：凝血致活酶　切入点：冠状病毒　出处：《中国病毒学(英文版)》2006年04期 　论文类型：期刊论文

【摘要】：研究鉴定激活hfgl2凝血酶原酶基因的SARS冠状病毒结构蛋白。从SARS尸检肺组织中抽提RNA后制备cDNA,分别扩增SARS-CoV的N、S2和M全长基因序列,再分别克隆到真核表达载体pcDNA3.1(+)上。应用免疫组织化学分析鉴定pcDNA3.1-N、pcDNA3.1-M和pcDNA3.1-S2的表达。构建人纤维介素(hfgl2)启动子荧光素酶报告基因质粒,并将SARS冠状病毒结构蛋白表达质粒分别与其共转染以明确激活hfgl2基因转录的SARS冠状病毒结构蛋白。将目的片段克隆至pcDNA3.1(+),经酶切鉴定和测序鉴定无误;免疫组织化学染色可见明显的CHO细胞胞浆棕染。与hfgl2启
[Abstract]:The structural protein of SARS coronavirus which activates the prothrombin gene of hfgl2 was identified. The RNA was extracted from the lung tissue of SARS autopsy to amplify the full-length sequence of SARS-CoV gene. Then the eukaryotic expression vector pcDNA3.1 () was cloned respectively. The expression of pcDNA3.1-M and pcDNA3.1-S2 was identified by immunohistochemical analysis. The luciferase reporter gene plasmid of human fibronectin hfgl2) promoter was constructed. The expression plasmid of SARS coronavirus structural protein was cotransfected with SARS coronavirus structural protein to activate the transcription of hfgl2 gene respectively. The target fragment was cloned into pcDNA3.1 and confirmed by restriction endonuclease digestion and sequencing. Immunohistochemical staining showed that the cytoplasm of CHO cells was brown stained. It was associated with hfgl2.
【作者单位】：华中科技大学同济医学院附属同济医院感染免疫研究室感染科南方医院感染内科华中科技大学同济医学院附属同济医院感染免疫研究室感染科南方医院感染内科华中科技大学同济医学院附属同济医院感染免疫研究室感染科华中科技大学同济医学院附属同济医院感染免疫研究室感染科
【基金】：科技部973-SARS专项(2003CB514112) 教育部首批SARS项目基金(教技司[2003]64号)
【分类号】：R373

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