一种改进的对虾白斑综合征病毒提纯方法  

A method of white spot syndrome virus (WSSV) purification

作　　者：姜有声[1,2] 战文斌[1] 程顺峰[1] 王世表[1] 

机构地区：[1]中国海洋大学教育部海水养殖重点实验室,山东青岛266003 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《上海海洋大学学报》2009年第3期372-375,共4页Journal of Shanghai Ocean University

基　　金：国家973课题基金(2006CB101806);上海高校选拔培养优秀青年教师科研专项基金项目(科07-05);上海海洋大学博士启动基金(科06-315)

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囊膜的病毒颗粒。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 （Carabarus 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 taillike envelope extension at one extremity. The nucleoeapsids 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.

关 键 词：白斑综合征病毒 病毒提纯 完整病毒 

分 类 号：S945.1[农业科学—水产养殖]