摘要
本文介绍一种对EHFV兼有灭活和裂解双重效用的分离方法,即低温变性凝胶过滤法,利用低温变性的原理,将病毒材料先于深低温-80℃冻融,继而于-30℃冻融多次,使其钝化与裂解,然后通过凝胶过滤,将残留的毒粒及其裂解产物包膜蛋白、RNA等分离出来.三者之间的得率受病毒材料体积大小、病毒浓度高低、以及培养液中有无小牛血清等制约.这是目前获取足量EHFV亚单位和RNA等最廉价的一种手段,
A purification procedure has been developed for the inactivation and split-ting of EHFV by means of the cryodenaturation and gel filtration method. which involves (1) freezing and thawing of virus material at -80℃,-30℃ and 8℃ respectively; (2) separation of the remaining EHFV and its splitting products (envelope proteins and RNAs) by gel filtration on Sephadex G-200: (3)their detection,which has revealed that the recovery rate varies with the volume of virus material,virus concentration and fetal calf serum in the culture. So far this method may proved to be the simplest and cheapest one for preparation of enough EHFV subunit and RNA.
关键词
出血热病毒
冻融
裂解
机理
Hemorrhagic fever virus
epidemic
Freezing
RNA
Protein denaturation
Protein degradation
