2002•Medical Journal of National Defending Forces in Northwest ChinaRequires access

Construction of eukaryotic expression plasmid for tumor suppression gene p27

Mao-long Dong

Abstract

Objective:To construct eukaryotic expression plasmid for tumor suppression gene p27 for further studying the effect of p27 on wound healing and scar forming.Methods:Synthesized complete cDNA by PCR, and cloned it into pUC19 for DNA sequence analysis; pUC19-p27 and pcDNA3 were digested with Hind III and EcoR I, then the p27 cDNA was lignated with pcDNA3 at Hind III and EcoR I restriction sites by T4 lignase and pcDNA3-p27 formed. Results:The synthesized p27 cDNA contained correct and complete DNA sequence ;p27 cDNA has been inserted into eukaryotic expression plasmid pcDNA3.Conclusion:The eukaryotic expression plasmid pcDNA3-p27 for tumor suppression gene P27 has been constructed successfully.

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Key concepts: pUC19, Plasmid, Complementary DNA, Molecular biology, Gene, Biology, Transfection, Gene expression