Construction of Plasmids Expressing Sars-CoV Encoding Proteins and Their Effects on Transcription of Hfgl2 Prothrombinase  

作　　者：王洪武 韩梅芳 姚华宁 王战会 习东 严伟明 侯金林 罗小平 宁琴 

机构地区：[1]Department of Infectious Disease,Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology [2]Department of Infectious Diseases, Nanfang Hospital, Nanfang Medical University [3]Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology

出　　处：《Journal of Huazhong University of Science and Technology(Medical Sciences)》2009年第3期318-323,共6页华中科技大学学报（医学英德文版）

基　　金：supported by a grant from National Key Project of Science and Technology Ministry of China for 973-SARS (No. 2003CB514112);SARS funding first granted from Ministry of education of China ([2003]64);The National 10th Five-Year Plan Key Project of China (2004BA720A01)

摘　　要：SARS coronavirus （SARS-CoV） is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane （M）, nucleocapsid （N） and spike 2 （$2） gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and $2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.SARS coronavirus （SARS-CoV） is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane （M）, nucleocapsid （N） and spike 2 （$2） gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and $2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.

关 键 词：SARS-COV encoding protein gene expression 

分 类 号：R373[医药卫生—病原生物学]