Objective To establish a eukaryotic cell line that can express soluble human leucocyte antigen G1(sHLA-G1) stably. Methods The recombinant plasmid pcDNA3-sHLA-G1 is transfected by a novel nonviral, electroporation-based gene transfer method termed nucleofection into the host cell lymphoblastoid cell line (LCL)721.221 which does not express any HLA-classical I molecules. After selection by G418, the cell line stably expressingsHLA-G1 is identified by RTPCR and Dot-ELISA with HLA-G1 specific monoclonal antibody MEM-G/9. Results The efficiency of transfection for LCL721.221 is about 14% by nucleofection. The specific band forsHLA-G1 was found by RT-PCR assay from the transfections and the protein ofsHLA-G1 in the supernatant of the transfections was detected by Dot-ELISA assay. Both confirmed that the eukaryotic cell line expressingsHLA-G1 has been established successfully at genic and proteinic levels. Conclusion In this study, the eukaryotic cell line expressingsHLA-G1 have been established successfully by nucleofection.
Citation: ZENG Menghua,FANG Chongyun,WANG Shusen,et al.. ESTABLISHMENT OF EUKARYOTIC CELL LINE STABLY EXPRESSING SOLUBLE HUMANLEUCOCYTE ANTIGEN G1 BY NUCLEOFECTION. Chinese Journal of Reparative and Reconstructive Surgery, 2006, 20(11): 1130-1133. doi: Copy