Construction and identification of the recombinant eukaryotic expression plasmid pHsp70-hsv-tk / 医学研究生学报

ABSTRACT

Objective: To construct the recombinant eukaryotic expression plasmid pHsp70-hsv-tk(pHT)for further target gene therapy research.Methods: The oligomers of primers were synthesized based on the reported sequences of hsv-tk and with the pHSV-106 plasmid as the template.Then the hsv-tk gene fragment was amplified and inserted into the pD3SX vector.E.coli DH5? was transformed with the recombinant plasmids and the bacterial colonies containing the recombinant plasmids were identified by the PCR cloning method,and the recombinant plasmids were extracted and purified.Results: The recombinant eukaryotic expression plasmid pHT was obtained.The target gene obtained by PCR amplification had the same molecular size as predicted.The recombined pD3SX plasmid contained correct recombinant hsv-tk sequences.Conclusion: The plasmid pHT was successfully constructed,which can be used for further target gene therapy research.