摘要
作者从发生白点综合征的中国对虾中，提取了一株白点综合征病毒，用电镜对纯化的病毒进行了形态学观察，并对病毒纯化过程中蛋白酶抑制剂的使用及离心速度的选择进行了探讨。经蔗糖密度梯度离心，分取各带负染后电镜观察，病毒条带位于４０％～５０％蔗糖梯度之间，其完整毒粒长２２５～２７０ｎｍ，直径７５～８８ｎｍ，有的完整毒粒带有很长的尾，病毒衣壳长２５０～３２０ｎｍ，直径６０～８０ｎｍ。在匀浆使用的缓冲液中加入蛋白酶抑制剂ＰＭＳＦ，可得到完整的带包膜的病毒颗粒，比不使用ＰＭＳＦ的提纯效果要好。经３０００ｒ／ｍｉｎ离心５ｍｉｎ，５０００ｒ／ｍｉｎ离心２５ｍｉｎ，去除大部分的组织碎片，经１００００ｒ／ｍｉｎ离心１ｈ，可使大部分病毒沉淀。并采用传统的方法对病毒核酸进行了提取，其基因组经初步分析为ｄｓＤＮＡ。
A stain of WSSV was isolated and purified from P.Chinensis . The morphology and structure of this virus was observed by transmissoin electron microscope (TEM). The procedures for the purification of virions and extraction of the viral nucleic acid of WSSV of shrimps are described. After centrifuged in sucrose gradient, each band was collected, negatively stained and examined by transmission electron microscope. The banded virus was located between 40%￣50% sucrose gradient. The length of enveloped virions was 225￣270nm and the diameter was 75￣88nm. The nucleocapsids were measured to be 250nm in length and 60￣80nm in width. The nucleic acid of virus was extracted by a general protocol and the genome was dsDNA by primary analysis.
基金
联合国教科文组织项目
关键词
白点综合征病毒
中国对虾
病毒核酸
病毒纯化
white spot syndrome virus(WSSV)
Penaeus Chinensis
viral nucleic acid
virus purification
Acknowledgements
