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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。
The WSSV genome was isolated by purifying the particles of WSSV, extracting the viral DNA, digesting it with restriction endonuclease EcoRI or SalI, and cloning into plasmid pBluescriptIIKS. The WSSV genomic DNA is estimated to be above 16 5 kb. The WSSVEcoRI cloning fragment marker was prepared as a probe for Southern hybridization, dot blot hybridization and in situ hybridization. The results confirmed that the cloned fragment was specific to WSSV and provided a method for detecting WSSV. The sequence analysis of partial genomic library revealed that the WSSV nucleic acid sequence has low homology with known baculovirus nucleic acid sequences and no significant homology with the existing virus nucleic acid sequences in GenBank, indicating that WSSV is not a baculovirus Family, currently only classified as unclassified invertebrate virus, the taxonomic status still needs more evidence to determine.