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  

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

作　　者：于萍[1] 刘瑾[1] 张立[1] 李胜富[1] 步宏[1] 李幼平[1] 程惊秋[1] 陆燕蓉[1] 龙丹[1] 

机构地区：[1]四川大学华西医院卫生部移植工程与移植免疫重点实验室,成都610041

出　　处：《四川大学学报（医学版）》2005年第6期770-772,共3页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30070725);纽约中华医学基金会(CMB00-722)资助

摘　　要：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(BM I)骨髓间充质干细胞(M SC s)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、po l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BM I-M SC s至连续传代到150、180代的BM I-M SC s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BM I近交过程中及猪细胞体外长期传代过程中均未消失,PERV基因已经整合入其天然宿主的基因组内并能稳定表达。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 PEP, V-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.

关 键 词：猪内源性逆转录病毒 版纳微型猪近交系 永生细胞系 异种器官移植 人兽共患病 

分 类 号：S852.65[农业科学—基础兽医学]