Résumé
To find out whether the integration of PERV in HEK293 cells influence the expression profile of HERV-W,based on the gene sequences of H ERV-W in GenBank,the primers of H ERV-W gag,pol,env,env from locus 7q21.2 genes as well as humanβ-actin were synthesized respectively and were used to develop the means of SYBR Green I real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to detect the mRNA expression of these genes in HEK293 cells and PERV-infected HEK293 cells.Experiments showed that these RT-qPCR methods were of good specificity,sensitivity and stability:the standard curves could ensure the correlation coefficients to be all above 0.99 and the amplification efficiency were between 95% and 110%,which verified that these methods could be used to detect the mRNA of HERV-W and humanβ-actin in culture cells.Through the detection and analysis of relative gene expression data using the 2-△△ct method,we found that the mRNA level of HERV-W gag,pol,env,env from locus 7q21.2 genes as well as humanβ-actin in PERV-infected HEK293 cells increased,after integration,by 37.08,42.56,2.49 and 13.17 times than in non-infected HEK293 cells,respectively.Results provide a reference to further evaluate the safety of PERV pathogen in xenotransplantation.