Abstract:

Abstract:Objective To construct the Cre rebombinase expression vector and identify its function of recognizing loxP sites and provide the basis for establishment of the animal models of human diseases. Methods The recombinant vectors Cre-pCEP4 and pStop-eGFP were constructed. The MCF-7 cells and porcine embryonic fibroblasts (PEF) were transfected by Fugene HD. The expression of eGFP was analyzed by fluorescence microscope. Results The recombinant vectors Cre-pCEP4 and pStop-eGFP were constructed successfully and they were co-transfected into PEF.The  MCF-7 cells with stable expression of Cre recombinase were transfected with pStop-eGFP after selection of hygromycin B. There were green fluorescent protein exrpression of eGFP analyzed by fluorescence microscope after transfected by the two vectors. No eGFP positive cell was detected in MCF-7 cells and PEF transfected with pStop-eGFP only. Conclusion The recombinant vector Cre-pCEP4 can express Cre recombinase which can recognize loxP sites and delete the DNA agment between two loxP sites in the same orientation.

Key words: Cre-pCEP4;Cre/loxP system;porcine embryonic fibroblasts;MCF-7 cell line