• Key Laborary of Organ Transplantation of Ministry of Education &;
  • Key Laborary of Organ Transplantation of Ministry of Public Health, Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei, 430030, P. R. China;
Soluble human leucocyte antigen G1; Nucleofection; Eukaryotic cell line
Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables References Cited by

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 RTPCR 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 G1 BY NUCLEOFECTION. Chinese Journal of Reparative and Reconstructive Surgery, 2006, 20(11): 1130-1133. doi: Copy