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2005•Journal of Jinan UniversityRequires access

Construction of eukaryotic plasmid including HCV NS3 gene and expression of NS3 protein in human hepatocytes

XU Xia-liang

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Abstract

Aim: To construct eukaryotic plasmid which includes HCV NS3 gene, and induce the expression of NS3 protein in human hepatocytes. Methods: HCV NS3 gene was cloned by using PCR from plasmid pBRTM/HCV1-3011 which includes the full length of HCV gene. We recombined the NS3 segment with expression plasmid pcDNA3.1(-) to construct eukaryotic plasmid pcDNA3.1(-)/NS3. Then plasmid pcDNA3.1(-)/NS3 was transfected into human hepatocytes by using poly-cation. The expression of NS3 protein was detected by immunohistochemical staining method and Western Blotting. Results: The cloned NS3 segment had correct length and sequence. The QSG7701 cells were successfully transfected and expressed NS3 protein. Conclusion: Eukaryotic expression plasmid pcDNA3.1(-)/NS3 incluing HCV NS3 gene was successfully constructed and expressed in human hepatocytes.

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What this paper is about

Aim: To construct eukaryotic plasmid which includes HCV NS3 gene, and induce the expression of NS3 protein in human hepatocytes. Methods: HCV NS3 gene was cloned by using PCR from plasmid pBRTM/HCV1-3011 which includes the full length of HCV gene. We recombined the NS3 segment with expression plasmid pcDNA3.1(-) to construct eukaryotic plasmid pcDNA3.1(-)/NS3. Then plasmid pcDNA3.1(-)/NS3 was transfected into human hepatocytes by using poly-cation. The expression of NS3 protein was detected by immunohistochemical staining method and Western Blotting. Results: The cloned NS3 segment had correct length and sequence. The QSG7701 cells were successfully transfected and expressed NS3 protein. Conclusion: Eukaryotic expression plasmid pcDNA3.1(-)/NS3 incluing HCV NS3 gene was successfully constructed and expressed in human hepatocytes.

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Available abstract

Aim: To construct eukaryotic plasmid which includes HCV NS3 gene, and induce the expression of NS3 protein in human hepatocytes. Methods: HCV NS3 gene was cloned by using PCR from plasmid pBRTM/HCV1-3011 which includes the full length of HCV gene. We recombined the NS3 segment with expression plasmid pcDNA3.1(-) to construct eukaryotic plasmid pcDNA3.1(-)/NS3. Then plasmid pcDNA3.1(-)/NS3 was transfected into human hepatocytes by using poly-cation. The expression of NS3 protein was detected by immunohistochemical staining method and Western Blotting. Results: The cloned NS3 segment had correct length and sequence. The QSG7701 cells were successfully transfected and expressed NS3 protein. Conclusion: Eukaryotic expression plasmid pcDNA3.1(-)/NS3 incluing HCV NS3 gene was successfully constructed and expressed in human hepatocytes.

Key concepts: Plasmid, NS3, Molecular biology, Transfection, Gene, Expression vector, Biology, Recombinant DNA

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