Cloning and expression of HLA-E cDNA on LcL721.221 cell

ObjectiveTo clone human HLA E cDNA and obtain stable expression on HLA classⅠnegative LcL721 221 cell Methods RT PCR technique was employed to amplify HLA E cDNA from human PBMC, the cDNA was subcloned into retrovirus vector pGCEN carrying HLA A2 cDNA through internal ribozyme entry site (IRES) thus the multi cistron expression vector was constructed and named pG/A2E Then infectious method was used to transduct the recombinant plasmid into the target cells followed by screening with G418 and limiting dilution; Finally, flow cytometry was adopted to detect HLA E expression on the target cells Results HLA E molecules were successfully expressed on LcL721 221 cells tranducted with pG/A2E (88 79%) and the expression was higher than control JAR (26 21%), but the expression of HLA E molecules was not detected on LcL721 221 cells transducted with pG/A2 Conclusion The multi cistron expression vector was constructed successfully and the HLA E molecules were expressed on LcL721 221 cells