Construction of Human CD244 Gene Transfected Cell Line

Objective To construct genetic transfecting cell line that stably expressed the human CD244.Methods Human CD244 gene was cloned by RT-PCR and subcloned into retroviral expressing vetor pEGZ-Term.The recombinant plasmid pEGZ-Term/CD244 together with its two helper virus vector were cotransfected into the package cell 293T with Lipofectamine 2000.The L929 cells were infected with the supernatant of the transfected 293T cells,and then were selected with Zeocin.The Zeocin resistant cells were harvested for screening their GFP and CD244 expression by fluorescence microscope and flowcytometry.Results Genetic transfecting line L929/CD244 which stably expressed the human CD244 was constructed.Conclusion Genetic transfecting cell line which stably expressed the human CD244 is established successfully.It provids a valuable tool to study the biological function of CD244 moelecule.