Construction of shRNA eukaryotic expression plasmid of MDR1 gene

To construct short hairpin RNA( shRNA) eukaryotic expression recombinant plasmid targeting MDR1 gene,three intervention targets were selected from the sequences corresponding to MDR1 gene,and were used to design and synthesize three DNA templates encoding shRNA. The shRNA templates were directionally cloned into the eukaryotic expression plasmid pSilencer 3. 1- H1 to construct the recombinant plasmid, respectively,and the shRNA recombinant plasmid pSilencer 3. 1- H1 were identified by colony- PCR amplification and DNA sequencing. The results showed that the positive bands same as the expected sizes were obtained by colony- PCR amplification. It was proved that all the inserted fragments were fully consistent with the design requirements,and there were no mutations in the sequences. The results indicate that the three the eukaryotic expression recombinant plasmids pSilencer 3. 1- H1 shRNA targeting MDR1 gene are successfully constructed.