Construction of Recombinant Plasmid Series with HSP70-2-Targeted Short Hairpin RNAs

Objective: To construct the recombinant plasmid expressing HSP70-2-targeted short hairpin RNA(shRNA) by pGenesil-1 plasmids vector.Methods: Three pairs of DNA sequences were designed,and then synthesized into complementary chains by annealing,respectively.Then the obtained products containing short hairpin structure were inserted into plasmid vector pGenesil-1 with U6 promoter.The recombinant plasmids were transformed into Escherichia coli strain DH5α for screening and amplifying.The sequence analysis of the plasmids was identified by restriction enzyme.Results: The three HSP70-2-targeted shRNAs were successfully inserted into the plasmid vector pGenesil-1,and the coding sequences of the obtained shRNAs were consistent with the designed fragments.pGenesil-1-HSP70-23 exerted the most significant inhibition effect on HSP70-2 gene.Conclusion: The recombinant plasmid series of HSP70-2-targeted shRNAs is successfully constructed in pGenesil1 plasmid.