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Construction of Recombinant Plasmid Containing Sna(2007-9-17 21:41:21) |
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Objective To construct recombinant plasmid of snake venom cystatin gene and its expression in the eukaryotic COS7 cells with the vector of pcDNA3.1/HisC. Methods Snake venom cystatin gene was amplified by PCR and inserted into pcDNA3.1/His C. The expression of COS7 cells was detected in by Westernblot assay. Results Cystatin gene was correctly inserted into the vector pcDNA3.1/His C, which was identified by the endonuclease digestion and sequencing. Cystatin can be detected in the plasmid pcDNA3.1/Hiscystatin transfected COS7 cells by Westernblot assay. Conclusion Eukaryotic expression plasmid pcDNA3.1/Hiscystatin was constructed successfully, identifying by it could be expressed in the COS7 cells.
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