猪内源性逆转录病毒基因在版纳微型猪近交系骨髓间充质干细胞永生细胞系的整合和表达研究

目的 检测猪内源性逆转录病毒（PERV）在猪细胞长期传代过程中的基因整合和表达。方法 通过提取已建立的版纳微型猪近交系（BMI）骨髓问充质干细胞（MSCs）永生细胞系的DNA及细胞总RNA，用PCR、RT—PCR方法检测PERV前病毒gag、pol和env基因的整合和表达，并对PERV的亚型进行鉴定。结果 从原代BMI—MSCs至连续传代到150、180代的BMI—MSCs细胞基因组中都检测到了PERV前病毒DNA的整合及其mRNA的表达．PERV亚型为PERV—A、B型。结论 PERV在BMI近交过程中及猪细胞体外长期传代过程中均未消扔，PFRV基因已经整合入其天然宿主的基因组内并能稳定表达。

Integration and Expression of Porcine Endogenous Retrovirus in the Immortal Cell Line of Banna Minipig Inbred Line-Mesenchymal Stem Cells

Objective To detect the integration and expression of porcine endogenous retrovirus(PERV) in the immortal cell line of Banna Minipig Inbred Line-Mesenchymal Stem Cells(BMI-MSCs).Methods DNA and total RNA of the immortal cell line of BMI-MSCs were extracted and PCR, RT-PCR were performed to detect PERV-gag,pol and env gene,and the type of PERV was also detected.Results PERV-gag,pol and env gene were all detected in the primary culture and immortal cell line(passage 150 and passage 180) of BMI-MSCs,and the type of PERV was PERV-A,B.Functional expression of PERV-gag and pol mRNA was also detected.Conclusion In this laboratory,PERV was not lost during the proceeding of pig inbred and since has been in long-term culture of pig cells in vitro.PERV has integrated into the genome of its natural host,and virus mRNA can effectively express.So it is very essential to evaluate the possibility of xenozoonoses in pig-to-human xenotransplantation.