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

• Published in: Journal of Huazhong University of Science and Technology. Medical sciences Vol. 29; no. 3; pp. 318 - 323
• Main Author: 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
• Format: Journal Article
• Language: English
• Published: Heidelberg Huazhong University of Science and Technology 01.06.2009; Department of Infectious Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China%Department of Infectious Diseases, Nanfang Hospital, Nanfang Medical University, Guangzhou 510515, China%Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
• Subjects: Animals; CHO Cells; Cricetinae; Cricetulus; Fibrinogen - biosynthesis; Fibrinogen - genetics; Genetic Vectors - genetics; Humans; Medicine; Medicine & Public Health; Membrane Glycoproteins - biosynthesis; Membrane Glycoproteins - genetics; Nucleocapsid Proteins - biosynthesis; Nucleocapsid Proteins - genetics; Plasmids; Recombinant Proteins - biosynthesis; SARS coronavirus; SARS Virus - genetics; SARS Virus - metabolism; SARS-CoV; Severe Acute Respiratory Syndrome - virology; Spike Glycoprotein, Coronavirus; Thromboplastin - genetics; Thromboplastin - metabolism; Transcription, Genetic - genetics; Transfection; Viral Envelope Proteins - biosynthesis; Viral Envelope Proteins - genetics; Viral Matrix Proteins - biosynthesis; Viral Matrix Proteins - genetics; 传染病; 感染病毒; 遗传基因; encoding protein; gene expression; SARS-CoV
• ISSN: 1672-0733; 1993-1352
• DOI: 10.1007/s11596-009-0311-1

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.