上海梅山猪封闭群内源性逆转录病毒的检测及亚型分析  

Investigation and Analysis of the Subtype of Porcine Endogenous Retrovirus in Breeding Base of Meishan Pigs in Shanghai

作　　者：吴鋆龙 张德福[2,3] 吴彩凤[2,3] 张树山[2,3] 王昕[1,2,3] 戴建军[2,3] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]上海市农业科学院畜牧兽医研究所,上海201106 [3]上海农业遗传育种重点实验室动物遗传工程研究室,上海201106

出　　处：《中国农学通报》2015年第14期39-43,共5页Chinese Agricultural Science Bulletin

基　　金：上海市科委科技兴农专项"克隆和冷冻保存技术在上海白猪保种;育种中的应用"(133919N1700);国家转基因生物新品种培育科技重大专项"不饱和脂肪酸;植酸酶转基因"环保猪"新品系的培育"(2009ZX08006-014B);"高繁殖力转基因猪新品种培育"(2014ZX08006-005);上海市农委青年人才成长计划"基于TALENs技术的猪外源基因定点整合方法的建立"(沪农青字2014-1-32);"甲基化修饰状态对猪体细胞克隆效率的影响"(沪农青字2014-1-33)

摘　　要：为检测分析上海梅山猪携带猪内源性逆转录病毒(porcine endogenous retrovirus,PERV)的情况,为该猪种作为器官移植供体的生物安全性提供现实依据,笔者从上海嘉定区梅山猪育种中心的梅山猪封闭群内随机采集45头猪的耳组织,并构建耳成纤维细胞系,利用PCR检测PERV前病毒核心蛋白基因(gag)、多聚酶基因(pol)以及囊膜基因(env)的基因组DNA,利用RT-PCR的方法检测m RNA表达状况。结果发现被检测的45份样品均携带完整的3种亚型的PERV前病毒DNA;RT-PCR结果显示,45份样品中有40份同时有PERV-A和PERV-B两种亚型的表达,还有5份样品仅检测到PERV-B亚型的表达,未检测到PERV-C亚型的表达。鉴于生物安全性方面的考虑,该猪种能否作为人异种移植的供体仍有待进一步商榷。In order to provide actual evidence of biological safety for using Meishan pigs for donor organtransplantation, existence and subtype of porcine endogenous retrovirus（PERV） in Meishan pigs wereinvestigated and analyzed respectively. Tissues of 45 pig ears were collected randomly from Meishan pigbreeding center in Jiading District, Shanghai. Ear fibroblast cell lines from these tissues were established. Theexistence of core protein gene（gag）, polymerase gene（pol） and capsule membrane gene（env） from PERVprovirus in genomic DNA was detected by PCR and expression of m RNA of each gene was tested by RT-PCR.The results showed that three subtypes PERV provirus existed in all 45 DNA samples. Results of RT-PCRshowed that PERV-A and PERV-B both expressed in 40 samples, subtype PERV- B expressed only in 5samples, and subtype PERV-C was not found in this study. In consideration of biological safety, whether thesepigs can be used as human xenotransplantation donors still need further discussion.

关 键 词：梅山猪 猪内源性逆转录病毒 病毒亚型 异种移植 

分 类 号：S828.2[农业科学—畜牧学]