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Virologica Sinica, 21 (4) : 319-323, 2006
Research Article
Nucleocapsid Protein of SARS-associated Coronavirus Upregulates the Transcription of hfgl2 Prothrombina -se/fibroleukin Gene
1.Department of Infectious Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
2.Department of Infectious Diseases, Nanfang Hospital, Guangzhou 510515, China
The structural protein of SARS-associated coronavirus (SARS-CoV) responsible for the activation of hfgl2 gene was investigated. Gene fragments encoding the nucleocapsid, the membrane, and spike proteins were amplified by RT-PCR, and they were cloned into the eukaryotic expression vector pcDNA3.1(+). The plasmids were verified by restriction endonuclease analysis and sequencing. Chinese hamster ovary (CHO) cells were co-transfected with a plasmid carrying the hfgl2 promoter/luciferase and with one of the recombinant plasmids containing the genes enconding the N, M and S structural proteins. Luciferase activity was assayed as a measure of promoter function. Immunohistochemistry showed that genes encoding the structural protein of SARS-CoV were successfully expressed only in those cells transfected with the recombinated plasmids. Co-transfection of N gene expression construct with the reporter construct containing hfgl2 promoter in CHO cells showed a remarkable increase in luciferase activity compared with nontransfected cells. However, there was no significant difference in luciferase activity in cells cotransfected with pcDNA3.1-M and with pcDNA3.1-S2 along with hfgl2 promoter/ luciferase reporter gene. We conclude that the nucleocapsid protein of SARS-CoV upregulates the transcription of hfgl2 prothrombinase/fibroleukin gene.
  Published online: 20 Jul 2006
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