Construction of eukaryotic expression vector for kainate receptor subunit GluR6 specific shRNA

Objective To construct the eukaryotic expression plasmid for kainate receptor(KA)subunit GluR6 specific shRNA.Methods According to the GluR6 cDNA sequence,two strands of oligonucleotide were designed and chemically synthesized,and then annealed into two double strands in vitro.The DNA strands were ligated with plasmid vector pSilencer-GluR6.The recombinate plasmid was transfected into E.coli and multiplied.After being extracted from E.coli,the plasmids were processed to verify the sequence of the shRNA by restriction endonuclease digestion and sequencing.Results The recombinant plasmid was found containing correct and complete sequence of the shRNA,indicating that the specific GluR6 shRNA had been inserted into the eukaryotic expression plasmid.Conclusion The eukaryotic expression plasmid pSilencer-GluR6 has been successfully constructed.