Construction of recombinant plasmid with RhoA-targeted short hairpin RNA

Objective To construct the recombinant plasmid expressing RhoA-targeted short hairpin RNA(shRNA)using pGenesil-1 plasmid vector.Methods Two pairs of DNA sequences were designed,and synthesized into complementary chains by annealing.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 DH5a for screening and amplification.The sequence analysis of the plasmids identified by restrictive enzyme techniques was carried out.Results The two RhoA-targeted shRNAs were successfully inserted into the plasmid vector pGenesil-1,and the coding sequences of the obtained shRNA were consistent with the designed fragments.Conclusion The recombinant plasmid of RhoA-targeted shRNA was successfully constructed.