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当前位置: 首页 > 文章 > 一种改进的对虾白斑综合征病毒提纯方法 上海海洋大学学报 2009,18 (3) 372-375
Position: Home > Articles > A method of white spot syndrome virus (WSSV) purification Journal of Shanghai Ocean University 2009,18 (3) 372-375

一种改进的对虾白斑综合征病毒提纯方法

作  者：
姜有声;战文斌;程顺峰;王世表
单  位：
上海海洋大学水产与生命学院;中国海洋大学海水养殖教育部重点实验室;连云港市海洋与水产科学研究所
关键词：
白斑综合征病毒;病毒提纯;完整病毒
摘  要：
对虾白斑综合征病毒(WSSV)的囊膜蛋白在提纯的过程中易脱落,用传统的密度梯度离心方法不易获得完整的病毒粒子,对传统的蔗糖密度梯度离心方法加以改进后,可以获得大量完整的病毒粒子。用患白斑综合征病毒病的中国对虾鳃制备WSSV粗提液,感染螯虾(Cambarus proclarkii),选取25%的蔗糖溶液用作病虾鳃的匀浆液,然后经蔗糖密度梯度离心,分取各带负染后电镜观察,大量完整的病毒位于46%~52%蔗糖梯度之间,病毒粒子末端带有很长的尾;而病毒裸露的核衣壳位于40%~46%蔗糖梯度之间;在57%~62%之间观察到较多完整病毒粒子和少量细菌。实验结果表明改进的病毒提纯方法较好,可得到大量完整的带嚢膜的病毒颗??
译  名：
A method of white spot syndrome virus (WSSV) purification
作  者：
JIANG You-sheng1,2,ZHAN Wen-bin1,CHENG Shun-feng1,WANG Shi-biao1(1.Laboratory of Pathology and Immunology of Aquatic Animals,LMMEC,Ocean University of China,Qingdao 266003,China;2.College of Fisheries and life Science,Shanghai Ocean University,Shanghai 201306,China)
关键词：
white spot syndrome virus;virus purification;intact virus
摘  要：
The envelope proteins of white spot syndrome virus(WSSV) are easy to be destroyed during purification.It was difficult to obtain intact virions by routine sucrose gradient centrifugation.After modifying the sucrose gradient,we can obtain a large quantity of intact virions.WSSV inoculums were prepared from gills of heavily infected Fenneropenaeus chinensis,and infected crayfish(Cambarus proclarkii),WSSV was purified from the gills of infected crayfish,25% sucrose was selected as homogenate medium.After being centrifuged in sucrose gradient,each band was collected and examined by transmission electron microscope.Enveloped virus was located between 46%-52% sucrose gradient,and most viral particles had a long tail-like envelope extension at one extremity.The nucleocapsids were between 40%-46% sucrose gradient.Between 57%-62% sucrose gradient,there were some bacteria and intact virus.The result showed this modified technique is very efficient for virus purification.

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