Effects of the integration of PERV on HERV-W mRNA expression in HEK293 cells / 中国人兽共患病学报

ABSTRACT

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 stabilitythe 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.