Cloning and expression of human leukocyte antigen-G mutant cDNA on K562 cell

ObjectiveTo obtain human HLA G mutant cDNA and stable e xpression on HLA class Ⅰ negative K562 cells. Methods RT P CR technique was used to amplify HLA G cDNA from human decidual tissue and the full length HLA G cDNA was obtained, and the product was point mutated by a pai r of specific primer, the mutant cDNA was subcloned into retrovirus vector pLNCX and the combined plasmid named mG pLNCX. Using infection technique to transduct the recombined plasmid into the target cells followed by screening with G418 and li miting dilution; Finally, flow cytometry was adopted to detect HLA G mutant exp ression on the cells.Results HLA G mutant molecule was success fully expressed on K562 cells transducted with mG pLNCX and no expression of H LA G on the control cell transducted with plasmid pLNCX.Conclusions the vector was contructed successfully and the HLA G mutant was expressed stably on the K562 cells.