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Construction and detection of the plasmid vector with the human U6 snRNA promoter for RNA Interference
Pages: 380-384,363
Year: Issue:  5
Journal: Journal of Zhenjiang Medical College

Keyword:  RNAiU6 promoterEGFPp53plasmid;
Abstract: Objective: To construct a vector to get the siRNAs for RNAi.Methods: The human U6 snRNA promoter from human genome was amplified by PCR,cloned into the reconstructed plasmid to generate the vector for RNAi,which called pUC19NU.To test the RNAi responses directed by the vector,designed two DNA templates that can be transcripted into shRNA against enhanced green fluorescent protein(EGFP) or p53 protein in cells and inserted them into pUC19NU respectively,then we got two vectors called pUC19NUE and pUC19NUP. Results: After being co-transfected into HeLa with pEGFP-N2 for negative contrast,the vector pUC19NUE was found to inhibit the EGFP gene almost completely.And after being transfected into KMB-17,the vector pUC19NUP also inhibit the p53 gene effectively. Conclusion: The vector we have constructed was efficient in arising RNAi response.Therefore,it can be used as a tool in future researches base on the technology of RNAi.
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