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[The expression and activity detection of a variant N protein of SARS-CoV].

Qi Yang, Yu Zheng, Cui-li Shu, Jiang Li, Yan Hu, Panyong Mao, Yun Cheng

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Abstract

AIM: To construct an expression vector pGEX-2T/N, and to express the fusion protein consisting of N protein of SARS-CoV and GST in E.coli. METHODS: The N region gene of SARS-CoV was cloned by RT-PCR. The expression vector was constructed by DNA recombination. The recombinant plasmid was transformed into E.coli BL21(DE3). The expression of the fusion protein was detected by Western blot. RESULTS: (1) As compared with the sequences in GenBank, 20 bp were deleted in DNA sequence of the cloned N protein. (2) The fusion protein GST-N was soluble. Western blot analysis showed that the reaction of GST-N to anti-SARS-CoV sera was positive. CONCLUSION: The pGEX-2T/N has been constructed and expressed in the form of fusion protein GST-N successfully, which lays the foundation for further study of SARS-CoV N protein.

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What this paper is about

AIM: To construct an expression vector pGEX-2T/N, and to express the fusion protein consisting of N protein of SARS-CoV and GST in E.coli. METHODS: The N region gene of SARS-CoV was cloned by RT-PCR. The expression vector was constructed by DNA recombination. The recombinant plasmid was transformed into E.coli BL21(DE3). The expression of the fusion protein was detected by Western blot. RESULTS: (1) As compared with the sequences in GenBank, 20 bp were deleted in DNA sequence of the cloned N protein. (2) The fusion protein GST-N was soluble. Western blot analysis showed that the reaction of GST-N to anti-SARS-CoV sera was positive. CONCLUSION: The pGEX-2T/N has been constructed and expressed in the form of fusion protein GST-N successfully, which lays the foundation for further study of SARS-CoV N protein.

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Available abstract

AIM: To construct an expression vector pGEX-2T/N, and to express the fusion protein consisting of N protein of SARS-CoV and GST in E.coli. METHODS: The N region gene of SARS-CoV was cloned by RT-PCR. The expression vector was constructed by DNA recombination. The recombinant plasmid was transformed into E.coli BL21(DE3). The expression of the fusion protein was detected by Western blot. RESULTS: (1) As compared with the sequences in GenBank, 20 bp were deleted in DNA sequence of the cloned N protein. (2) The fusion protein GST-N was soluble. Western blot analysis showed that the reaction of GST-N to anti-SARS-CoV sera was positive. CONCLUSION: The pGEX-2T/N has been constructed and expressed in the form of fusion protein GST-N successfully, which lays the foundation for further study of SARS-CoV N protein.

Key concepts: Fusion protein, Molecular biology, Recombinant DNA, GenBank, Western blot, Expression vector, Plasmid, Gene

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