Expression and identification of human EpCAM eukaryotic recombinant plasmid / 国际检验医学杂志

ABSTRACT

Objective To construct human EpCAM eukaryotic recombinant plasmid ,and to study the expression of EpCAM pro‐tein in HepG2 cells .Methods The recombinant plasmid ,named pcDNA3 .1(+ )‐EpCAMwas constructed by cloning EpCAM gene into eukaryotic vector pcDNA3 .1(+ ) .Then HepG2 cells were transfected with EpCAMrecombinant plasmid .The eukaryotic ex‐pression of EpCAM protein was verified by immunofluorescence ,Western Blotting and flow cytometry .Results The construction of EpCAM recombinant eukaryotic plasmid was identified by restriction enzyme digestion .The results of immunofluorescence , Western Blotting ,and flow cytometry consistently indicated that EpCAM protein were successfully expressed in HepG2 cells .Con‐clusion Human EpCAM eukaryotic vector is constructed and EpCAM protein could be expressed well in HepG2 cells ,which laid a foundation for further research on the function ofhepatocarcinoma cells highly expressing EpCAM .