2011•Xinxiang yixueyuan xuebaoRequires access
Construction and screening of short hairpin RNA recombinant plasmid expression vector targeting transketolase
Huigen Feng
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Abstract
Objective To screen the recombinant plasmid expressing TK-targeted short hairpin RNA(shRNA).Methods Aimed at TK gene order,four pairs of DNA sequences(hMGFP-shRNA1,hMGFP-shRNA2,hMGFP-shRNA3,hMGFP-shRNA4) were designed.The four recombinant plasmid were identified by PstⅠrestriction enzyme.Then the four plasmids were transfected into mouse hepatoma cell line Hepa1-6 for shRNA screening by reverse transcription-polymerase chain reaction and TK enzyme activity analysis.Results The four shRNA recombinant plasmids were correct verifying by PstⅠrestriction enzyme.shRNA screening identified that hMGFP-shRNA1 exerted the most significant inhibition effect on TK gene.Conclusion The recombinant plasmid of TK-targeted shRNAs is successfully screened.