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鸭瘟病毒的纯化及电镜负染形态观察
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郭宇飞1，2，程安春1，2*，汪铭书1，2，周毅2
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(1．四川农业大学 动物医学院 禽病防治研究中心，四川 雅安625014；
2．动物疫病与人类健康四川省重点实验室，四川 雅安625014)
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摘要：
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先做切向超滤浓缩，再采用蔗糖垫底超速离心和蔗糖密度梯度超速离心的方法对鸭胚成纤维细胞培养物中的鸭瘟病毒进行纯化，并对纯化的病毒粒子用电镜进行了观察。结果显示，用切向超滤法将病毒培养物浓缩为原体积的1/5后，再采用300g/L蔗糖垫底超速离心和300～600g/kg蔗糖密度梯度离心的方法进行纯化，可获得大量的纯化病毒粒子。电镜负染观察结果表明，病毒粒子呈圆形，多数直径为150nm左右，只有1个核衣壳，少数病毒粒子直径可达300nm，有2个或多个核衣壳。 |
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关键词：
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鸭瘟病毒；鸭胚成纤维细胞；纯化；负染 |
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中图分类号：S 852.659.1 文献标识码：
A 文章编号：1673-4696(2008)05-0393-04 |
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Purification and electron microscopy observation
after negative staining of duck plague virus |
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GUO Yu-fei1,2,CHENG An-chun1,2,WANG
Ming-shu1,2,ZHOU Yi2 |
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(1.Research Center for Avian Diseases,College of Veterinary Medicine，Sichuan Agricultural University,Ya’an
625014,China;2.Key Laboratory of Animal Diseases and Human Health of Sichuan Province,Ya’an 625014,China) |
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Abstract: Duck plague virus(DPV) in duck embryo fibroblast culture was purified sequentially by ultrafiltration,sucrose cushion ultracentrifugation and sucrose density gradient centrifugation.The morphology of the purified DPV virion was also observed by electron microscopy.The results showed that plenty of the purified DPV virions were obtained after the cell lysate was condensed by 5-fold by ultrafiltration,and the concentrated DPV fluid was ultracentrifuged with 300g/L sucrose cushion method and 300-600g/kg sucrose density gradient centrifugation methods.The purified DPV virions were round in shape with a diameter of approximately 150nm under electron microscopy.Most of the mature DPV virions had only one nucleocapsid and some of the mature DPV virions with a diameter of 300nm or more had two or more nucleocapsids.
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Key words: duck plague virus;duck embryo fibroblasts;purification;negative staining |
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