PERV在猪外周血白细胞DNA和组织mRNA中的表达及其差异性分析  

ANALYSIS OF PRESENCE AND DIFFERENCE OF SEQUENCE OF PORCINE ENDOGENOUS RETROVIRUS IN DNA GENOMES FROM PERIPHERAL BLOOD WHITE CELLS OF PIG AND mRNA IN TISSUES FROM MINI-PIGS

作　　者：丁生财[1] 陈意生[1] 魏泓[2] 商海涛[2] 刘萍[3] 

机构地区：[1]第三军医大学西南医院病理学研究所 [2]第三军医大学实验动物中心,重庆400038 [3]第三军医大学新桥医院妇产科,重庆400037

出　　处：《实验生物学报》2004年第5期351-358,共8页Acta Biologiae Experimentalis Sinica

基　　金：国家重点基础研究发展规划项目资助课题(No.G2000016106)。

摘　　要：为了解我国家猪猪内源性逆转录病毒(PERV)生物学的基本特征,为评价应用猪器官、组织、细胞进行猪—人间跨种移植的生物安全性提供理论基础。本文采用PCR方法调查12个家猪品系外周血白细胞DNA基因组PERV的生物学特征,并应用SS-SSCP、RFLP-PCR方法分析PERV基因片段的差异性及采用RT-PCR方法和半定量方法分析2个品系小型猪13种组织PERV表达的差异。结果表明12个品系猪外周血白细胞DNA基因组普遍存在PERV-A、-B基因序列,未发现单链构象多态性;部分品系猪PERV env基因序列片段存在限制性片段长度多态性。分析2个品系13种组织均表达PERV-A、-B、-C,肾、淋巴结、肝为高表达器官,胰腺和脑组织为低表达器官,PERV-C mRNA丰度明显低于PERV-A、-B mRNA。PERV env存在限制性片段长度多态性、PERV-A存在碱基缺失和错配的现象,有可能在猪异种移植中构成PERV感染的潜在危险性,这是在猪异种移植过程中值得高度关注的问题。The aim is to investigate biological features of porcine endogenous retrovirus (PERV)of pigs in China and to provide basic parameters for evaluation of biological safety of xenotransplantation from pig to human.In this study,basic biologic features of PERV of DNA of peripheral blood white cells of 12 species of domestic pigs in China were examined by poly- merase chain reaction;Analysis of difference of PERV gene sequence was investigated by sliver- stained single stranded conformational polymorphism and restriction fragment length polymorphism; Difference of PERV mRNA from a variety of tissue of two species of mini-pig was analysized by reverse transcriptase-PCR and semi-quantitative analysis.These results showed that Sequences of PERV-A,-B genes are ubiquitous in DNA genome of peripheral blood white cells in the exam- ined pigs;There was no single stranded conformational polymorphism in PERV-A,-B gene se- quences.There is the phenomenon of restriction fragment length polymorphism of sequence of PERV env gene in DNA genome of peripheral blood white cells in some pigs of the examined pigs.The PERV-A,-B and -C are expressive in a variety of tissues from two species of mini-pig, higher in kidneys,lymph nodes and livers,lower in pancreas and brains.It is deserved that the phenomenon of restriction fragment length polymorphism of PERV and the absence and mismatch of base of sequence of PERV-A may induce potential hazard of infection of PERV during xeno- transplantation.

关 键 词：外周血白细胞 低表达 异种移植 RNA 限制性片段长度多态性 错配 猪器官 品系 DNA基因 家猪 

分 类 号：S858.28[农业科学—临床兽医学]