聚乙二醇沉淀法部分提纯香石竹斑驳病毒  

PARTIAL PURIFICATION OF CARNATION MOTTLE VIRUS BY POLYETHYLENE GLYCOL PRECIPITATION

作　　者：张爱平[1] 

机构地区：[1]上海市园林科学研究所,上海200232

出　　处：《上海农业学报》1992年第2期88-90,共3页Acta Agriculturae Shanghai

基　　金：上海青年科学基金

摘　　要：接种香石竹斑驳病毒四周后的高雪轮病叶用2倍体积磷酸缓冲液匀浆。滤液用氯仿和正丁醇澄清,二轮聚乙二醇(PEG)沉淀,得到具有侵染活性的部分提纯的香石竹斑驳病毒。提纯制剂的A260/280值为1.612,病毒含量5.9mg/ml。与等量福氏不完全佐剂乳化后,4次肌肉注射家兔,得到双扩散效价为1:2048的专化抗血清。结果表明,PEG沉淀结合高速离心,是部分提纯香石竹斑驳病毒的可行方法。CV-68, an isolate of carnation mottle virus (CarMV) cultured in Silene armeria plants was harvested 4 weeks after inoculation and ground in a blender in two volumes of 0.5 mol phosphate buffer pH 7.2 containing 0. 01 mol EDTA-Na, 0. 01 mol ascorbie acid, 0.1% thioglycollic acid, 10% sucrose. The extract, yielded by straining through nyloneloth, was emulsified in 25% chloroform and 8% n-butanol (v/v), and clarified by low-speed eentrifugation. Then, two cycles of PEG precipiation (10% PEG, MW6000,3% NaCl) were used for virus separation from plant materials. Finally, a partially purified virus prepartion with high infectivity was obtained. The A260/280 ab sorption ratio of the preparation is 1. 612. In order to get specific antiserum against CV-68, one rabbit was immunized by four intramuscular injections of purified preparation emulsified in equal volume of Freund's incomplete adjuvant at weekly intervals. One week after the last injection, bleeding was taken and the antiserum with titre 1:2048 in Ouehterlony double diffusion test was collected, and mixed with glycerol, stored at —8℃——10℃.

关 键 词：香石竹 斑驳病毒 PEG沉淀 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]