猪细小病毒的提纯及其生化特性分析

Purification of Porcine Parvovirus and Analysis on its Biochemical Characteristics:

将自行分离鉴定的猪细小病毒 HZ-9株感染仔猪肾原代细胞,采用 PEG 沉淀法及 CsCl密度梯度离心从病毒细胞培养物中提纯病莓。该病毒 CsCl 梯度中获得单一的血凝峰,浮力密度为1.38～1.39g/ml,紫外扫描获得典型的病毒吸收峰,OD260mn/OD280nm 为1.64,电镜观察为球形颗粒,无囊膜,测得病毒粒子直径为19.6±1.8nm。经 SDS-PAGE 分析,病毒有3种结构多肽,其分子量和百分含量分别为 Vpl 81.7kd、9.8%、Vp2 67.6kd、11.7%,Vp364.6kd、76.9%。病毒核酸经中性1%琼脂糖凝胶电泳呈现一条核酸带,经碱性1%琼脂糖凝胶电泳测得核酸长度约为5kd,分子量约为1.44×10~6道尔顿。

The porcine parvovirus(PPV)HZ-9 strain isolated in the study was purified from infected porkling kidney cell cultures by high speed centrifugation with polyethylene glycol precipitation and CsCL quilibrirm density centrifugation.The virion particles possessing PPV specific hemagglutinating ac- tivity were banded at CsCL buoyant density of 1.38-1.39g/ml and were somestric,nonenveloped and icosahedral symmetric with a mean diameter of 19.6±1.8nm.The ultravirolet absorption spectra of puri- fied virus preparation was of typical virus,and the OD260/OD280 ratio was 1.64.The biochemical characteristics of PPV HZ-9 strain was partially studied.Analysis of Sodium Dodecyl Sulfat-Polyacrylamide Gel Electrophoresis(SDS-PAGE)demonstrated that the virion particles con- tained three major polypeptides Vp1,VP2 and Vp3 with molecular weight of 81.7Kd,67.6kd and 64.6kd respectively.The amount of polypeptides Vp1,Vp2 and Vp3 were 9.7%,11.7% and 76.9% respectively.The alkaline agarose gel electrophoresis indicated that viral nucleic acid was a single strand DNA with the length of 5 kd and the molecular weight of 1.44×10~6daltons.