作　　者： ; ;

机构地区： 中山大学生命科学学院

出　　处： 《水产学报》 2000年第2期161-166,共6页

摘　　要： 通过分离纯化白斑综合症病毒 (WSSV)粒子 ,抽提病毒DNA ,用限制性内切酶EcoRⅠ或SalⅠ酶切后 ,克隆入质粒pBluescriptⅡKS中 ,从而建立了WSSV部分基因组文库。估计WSSV基因组DNA在16 5kb以上。将WSSVEcoRⅠ克隆片段标记制备为探针 ,进行Southern杂交、打点杂交和原位杂交 ,其结果证明了克隆片段对WSSV特异 ,并为检测WSSV提供了方法。通过对部分基因组文库序列分析发现 ,WSSV核酸序列与已知杆状病毒核酸序列的同源性低 ,与GenBank中已有病毒核酸序列比较都无较大同源性 ,说明WSSV不属于杆状病毒科 ,目前只能列为未分类的无脊椎动物病毒 ,其分类地位尚需更多证据方能确定。 White spot syndrome virus(WSSV), a non-occluded virus which is the causative agent of a disease caused high shrimp mortalities and severe damage to shrimp cultures, was isolated and purified. Virions are rod-shaped, enveloped, 430nm×120nm in size, with a tail-like appendage at one extremity. Nucleocapsids are 300nm×85nm in size, and are composed of rings of subunits in a stacked series. The genome of the virus is a double-stranded DNA molecule. The full length of the DNA is estimated to be longer than 165kb. EcoRⅠ or SalⅠ restriction fragents obtained after agarose gel electrophoreses were cloned in the system pBluescript ⅡKS-XL1-blue. Southern hybridization, dot blot hybridization and in situ hybridization were performed using the WSSV EcoR Ⅰ clone fragments as probe, and those proved that these clone fragments are specific to WSSV. According to the results of nucleotide sequence analysis, none of the WSSV clones showed considerable sequence homology with those of other known viruses or organisms, indicating that WSSV may only be distantly related to baculoviruses.

关 键 词： 斑节对虾 白斑综合症病毒 核酸探针 基因组文库

领　　域： [农业科学] [农业科学]