摘要
采用氯仿、聚乙二醇-硫酸葡聚糖钠盐二相系统和蔗糖密度梯度离心法,从患病兔肝组织中提取、纯化病毒。该病毒粒子无囊膜,呈20面体对称,直径一般为33—37 nm。其三角剖分数 T=3,共有32个子粒,每一子粒为中空的、外径为9nm 左右的圆形轮廓。经蔗糖密度梯度离心后,可获得沉降系数为166s 的病毒颗粒,这种颗粒具有很强的感染性。经 SDS-PAGE 测得病毒粒子有四种多肽,分子量分别为66.4k、65.0k、63.5k、41.0k。二苯胺试验,吖啶橙染色、甲醛试验及热变性曲线表明,该病毒是单链 DNA 病毒。电镜下经甲酰胺法展层的病毒核酸呈单链线状,分子量平均为2.1×10~4道尔顿。此病毒的上述性质类似于细小病毒。53s 中空的圆形子粒,有很强的血凝性,而对照组铁蛋白和同一条件下所制备正常兔肝成份无凝血特性,说明53s 的9nm 粒子应为病毒外壳蛋白子粒。
In this paper a strain of Rabbit Hemorrhagic Disease Virus(RHDV)was isolated and pu-rified from the diseased rabbit livers with a method of using chloroform,two-phase of polyethy-lene-glycol-dextran sulfate sodium and sucrose density gradient centrifugation.Purified viruswas nonenveloped,icosahedeal symmetry with a triangulation number of 3,and 33—37nm indiameter.The capsid was composed of 32 capsomeres with central holes in an outer diameter ofabout 9nm.Two types of viral particles having different sedimentation coefficient,130s and166s could be identified after sucrose density gradient centrifugation.Probably no less than fourvirion proteins with molecular weight of 66.4,65.0,63.5,41.0×10~3 dalton were detected bySDS-polyacrylamide gel electrophoresis.Viral nucleic acid was extracted from purifiel virus byusing SDS-proteinase K-phenol.Tests with diphenylamine,formaldehyde,and staining withacridine orange as well as the curves of thermal denaturation showed that this kind of virushad a single-stranded DNA.The molecular weight of the ssDNA was approx 2.1×10~6 dalton asdetermined by electron microscopy.Data indicate that the RHDV may like the parvovirus of the family Parvoviridae.
出处
《微生物学报》
CAS
CSCD
北大核心
1992年第3期198-203,共6页
Acta Microbiologica Sinica
