猪生殖-呼吸道综合征国内分离毒株的病毒纯化及其结构蛋白分析  

Purification and Analysis of Major Structural Protein of the Porcine Reproductive and Respiratory Syndrome Virus Isolated in China

作　　者：蔡雪晖[1] 刘文兴[1] 郭宝清[1] 李成[1] 谷守林[1] 柴文君[1] 徐哮[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所,哈尔滨150001

出　　处：《中国预防兽医学报》1999年第4期272-274,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：中华农业科教基金;中国农业科学院国际合作基金

摘　　要：本研究主要对猪生殖呼吸道综合征（ Ｐ Ｒ Ｒ Ｓ） 国内分离毒株（ Ｃ Ｈ Ｉａ） 进行了病毒纯化及其结构蛋白的分析。选用聚乙二醇（ Ｐ Ｅ Ｇ６０００） 沉淀和差速离心法粗提了经 Ｍａｒｃ１４５ 细胞系增殖的猪生殖呼吸道综合征病毒（ Ｐ Ｒ Ｒ Ｓ Ｖ） 。并采用非线性蔗糖密度梯度离心法纯化了 Ｐ Ｒ Ｒ Ｓ Ｖ， 经免疫电镜观察纯化病毒， 发现有较多的胶体金颗粒吸附在直径为４２ ～７８ｎｍ 的病毒粒子周围。经 Ｄｏｔ Ｅ Ｌ Ｉ Ｓ Ａ 测定纯化病毒的滴度可达１ １６００ 以上， 并利用 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 和 Ｗｅｓｔｅｒｎｂｌｏｔ 两种方法对国内分离毒株（ Ｃ Ｈ Ｉａ） 与欧洲型（ Ｌｅｌｙｓｔａｄ） 、美洲型（ Ｖ Ｒ２３３２） 等参考毒株的病毒结构蛋白组成和部分抗原特性进行了初步的研究。测得国内分离毒株主要结构蛋白的分子量为１４５ Ｋ Ｄ，１９２ Ｋ Ｄ 和２６３ Ｋ Ｄ， 并均能被 Ｐ Ｒ Ｒ Ｓ Ｖ 的高免血清识别， 这与国外的同类报道很相似。In this study,porcine reproduetive and respiratory syndrome virus(PRRSV)isolaed in china was purified and its major structural protein were analyzed.PEG 6000,differential contrifugation and discontinuous sucrose density gradient centrifugation were adopred to concentrate and purify PRRSV from infected Marc 145 cells.Immune electron microscopy was used for detection of purified PRRSV,Sucrose purified virions visualzied by electron microscopy were spherial particles with an diameter range from 42nm to 78nm.And purified virions were surrounded by a number immuno gold particles,Purified virus was tested by Dot ELISA and its titer was≥11600.Antigencity and composition of PRRSV major structural proteins isolated in china was compared with those of European(Lelystad)and American type(VR 2332)by utilizing SDS PAGE and western blot.The 14.5 、19.3 and 26.3KD major structural protein of PRRSV have been determinded and were recognized by hyper immue anti PRRSV Sera.

关 键 词：PRRS 猪 生殖呼吸综合征 病毒 纯化 

分 类 号：S858.285.3[农业科学—临床兽医学] S852.65[农业科学—兽医学]