桡足类浮游动物细胞膜与白斑综合症病毒的结合研究  

Binding research between cell membrane of copepod and WSSV

作　　者：冯书营[1] 黄倢[1] 张士璀[2] 

机构地区：[1]中国水产科学研究院黄海水产研究所 [2]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《中国水产科学》2005年第4期458-464,共7页Journal of Fishery Sciences of China

基　　金：国家重点基础研究发展规划项目课题(G1999012002);国家自然科学基金课题(30271020);青岛市自然科学基金课题资助(032JZP10).~~

摘　　要：通过差速离心法提取桡足类组织细胞膜,依照本实验室确立的细胞膜与WSSV特异性的结合关系,进行了桡足类细胞膜与白斑综合症病毒(WSSV)结合实验。结果表明,桡足类细胞膜与WSSV之间存在着特异性的结合,二者的结合说明了桡足类细胞膜上存在有WSSV的受体蛋白,其为确定桡足类属于WSSV感染宿主提供了重要证据。同时,比较了凡纳滨对虾(Litopenaeusvannamei)鳃细胞膜对桡足类细胞膜与WSSV结合的影响作用,证实了凡纳滨对虾鳃细胞膜对二者的结合具有明显的封闭作用,侧面说明了WSSV在二者细胞膜上存在有相同的受体蛋白或结合位点。桡足类细胞膜经表面活性剂TritonX100处理后,与WSSV的结合活性没有受到影响,而经Tween20处理能显著提高与WSSV的结合活性,提示受体蛋白在细胞膜上可能占有一段较小的跨膜区域。经SDSPAGE分析确定了桡足类细胞膜由分子量为18～207kD范围的约33条蛋白条带组成。Infection host of White Spot Syndrome Virus (WSSV) is very wildly, such as Litopenaeus vannamei, Procambarus clarkii and Marsupenaeus japonicus, etc., but whether copepod belongs to the infection host above is still controversial. This paper focuses on investigating this debate through binding study between WSSV and cell membrane of copepod, which not only offers the evidence that copepod is of the infection host of WSSV, but also is quite valuable to further cognize the receptor of WSSV. Cell membrane from copepod was obtained through centrifugation and then was binded to the WSSV resorted to the specific binding relation which our Lab established between WSSV and cell membrane. The assay results showed that WSSV and cell membrane of copepod had a specific binding, this binding indicated receptor of WSSV on the cell membrane of copepod, it was a important proof to point that copepod belongs to the hosts of WSSV infection. This experiment also compared the influence of gill cell membrane of shrimp to the binding between WSSV and cell membrane of copepod and found that shrimp gill cell membrane had a distinctly close rolling to their binding. To some extent, this result proved that gill cell membrane of shrimp and copepod had the same WSSV binding location or receptor. Cell membrane of copepod was treated by triton X-100 and Tween 20 and then binded to WSSV, Tween 20 can enhance their binding but Triton X-100 can not. Cell membrane of copepod consist of 33 bands which MW range from 18?kD to 207?kD by SDS-PAGE analysis.

关 键 词：桡足类 细胞膜 WSSV 结合研究 

分 类 号：S941[农业科学—水产养殖]