Summary
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 (S2) gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and S2 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.
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Hongwu WANG and Meifang HAN contributed equally to the work.
This project was 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), and The National 10th Five-Year Plan Key Project of China (2004BA720A01).
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Wang, H., Han, M., Yao, H. et al. Construction of plasmids expressing Sars-CoV encoding proteins and their effects on transcription of hfgl2 prothrombinase. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 29, 318–323 (2009). https://doi.org/10.1007/s11596-009-0311-1
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DOI: https://doi.org/10.1007/s11596-009-0311-1