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Construction and identification of the siRNA expression vector of rat iNOS gene

Huang Jing-jing

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Abstract

Objective: To construct the siRNA expression vector of rat iNOS gene,which would be the base of establishing a platform of RNA interference technique. Methods: Using software to design the template of siRNA for rat iNOS gene. After synthesizing and annealling, it was inserted into pSilencerTM1.0-U6 vector to form a new clone——rat iNOS gene siRNA expression system. Then the recombinant plasmid was transformed into DH5α strain to amplify. The plasmid identified by restriction endounclease was used for sequence analysis after extraction. Results: Identifcation by enzyme cutting and sequencing showed that the expression vector was constructed successfully. Conclusion: The construction of the recombinant plasmid expressing siRNA of rat iNOS gene make the foundation for RNA interference technique, which will enable us to develop an approach on studying the mechanism and new therapy method for disease.

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What this paper is about

Objective: To construct the siRNA expression vector of rat iNOS gene,which would be the base of establishing a platform of RNA interference technique. Methods: Using software to design the template of siRNA for rat iNOS gene. After synthesizing and annealling, it was inserted into pSilencerTM1.0-U6 vector to form a new clone——rat iNOS gene siRNA expression system. Then the recombinant plasmid was transformed into DH5α strain to amplify. The plasmid identified by restriction endounclease was used for sequence analysis after extraction. Results: Identifcation by enzyme cutting and sequencing showed that the expression vector was constructed successfully. Conclusion: The construction of the recombinant plasmid expressing siRNA of rat iNOS gene make the foundation for RNA interference technique, which will enable us to develop an approach on studying the mechanism and new therapy method for disease.

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Available abstract

Objective: To construct the siRNA expression vector of rat iNOS gene,which would be the base of establishing a platform of RNA interference technique. Methods: Using software to design the template of siRNA for rat iNOS gene. After synthesizing and annealling, it was inserted into pSilencerTM1.0-U6 vector to form a new clone——rat iNOS gene siRNA expression system. Then the recombinant plasmid was transformed into DH5α strain to amplify. The plasmid identified by restriction endounclease was used for sequence analysis after extraction. Results: Identifcation by enzyme cutting and sequencing showed that the expression vector was constructed successfully. Conclusion: The construction of the recombinant plasmid expressing siRNA of rat iNOS gene make the foundation for RNA interference technique, which will enable us to develop an approach on studying the mechanism and new therapy method for disease.

Key concepts: RNA interference, Plasmid, Gene, Biology, Vector (molecular biology), Recombinant DNA, Molecular biology, clone (Java method)

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