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Construction eukaryotic expression vector of MCP-1 siRNA

Wang Shen-min

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Abstract

Objective To construct the small interfering RNA(siRNA) expression vector for monocyte chemotactite protein-1(MCP-1) and provide ways for artery stenosis treatment. Methods Two DNA sequences containing the sites of restriction endonuclease at both ends were designed and synthesized.The eomplement form was obtained by annealing and being cloned into vector pSilencer2.0-U6 and the recombinant plasmid was transformed into strain DH-5α. The plasmid identified by restriction enzyme was used for sequencing. Results MCP-1 siRNA expression vector was successfully constructed and identified by double endonuclease digestion.Sequence analysis of the inserted fragment revealed the salne sequence as that of the synthesized siRNA oligonucleotides. Conclusion MCP-1 siRNA expression vector has been successfully constructed, which lays the basis for its application in the prevention and treatment of artery stenosis.

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What this paper is about

Objective To construct the small interfering RNA(siRNA) expression vector for monocyte chemotactite protein-1(MCP-1) and provide ways for artery stenosis treatment. Methods Two DNA sequences containing the sites of restriction endonuclease at both ends were designed and synthesized.The eomplement form was obtained by annealing and being cloned into vector pSilencer2.0-U6 and the recombinant plasmid was transformed into strain DH-5α. The plasmid identified by restriction enzyme was used for sequencing. Results MCP-1 siRNA expression vector was successfully constructed and identified by double endonuclease digestion.Sequence analysis of the inserted fragment revealed the salne sequence as that of the synthesized siRNA oligonucleotides. Conclusion MCP-1 siRNA expression vector has been successfully constructed, which lays the basis for its application in the prevention and treatment of artery stenosis.

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Available abstract

Objective To construct the small interfering RNA(siRNA) expression vector for monocyte chemotactite protein-1(MCP-1) and provide ways for artery stenosis treatment. Methods Two DNA sequences containing the sites of restriction endonuclease at both ends were designed and synthesized.The eomplement form was obtained by annealing and being cloned into vector pSilencer2.0-U6 and the recombinant plasmid was transformed into strain DH-5α. The plasmid identified by restriction enzyme was used for sequencing. Results MCP-1 siRNA expression vector was successfully constructed and identified by double endonuclease digestion.Sequence analysis of the inserted fragment revealed the salne sequence as that of the synthesized siRNA oligonucleotides. Conclusion MCP-1 siRNA expression vector has been successfully constructed, which lays the basis for its application in the prevention and treatment of artery stenosis.

Key concepts: Restriction enzyme, Oligonucleotide, Molecular biology, Small interfering RNA, Plasmid, Recombinant DNA, Vector (molecular biology), Expression vector

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