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根据GenBank中登录的对虾白斑综合症病毒(WSSV)全基因组序列中和ORF234相应的序列来设计1对引物,从疑似病例的罗氏沼虾中提取总DNA,并以此为模板,经PCR扩增出约885bp的特异性片段,克隆进质粒载体pET-28a(+),进行序列测定和分析。结果显示:扩增序列共编码294个氨基酸,预测的相对分子质量为34kDa,与GenBank中登录序列的对应区域同源性达99%,由此确认该罗氏沼虾患有白斑综合病毒病。该序列所编码的蛋白在22～96氨基酸位置有一段球状结构,在142～272氨基酸位置有一个与DUF1335蛋白同源的将近130个氨基酸残基的保守区,其功能未知。同时还对克隆出的片段进行序列分析和蛋白预测,以进一步深入开展蛋白的功能及对该病毒病的防治的研究。
One pair of primers was designed according to the sequence of the whole genome of WSSV registered in GenBank and the corresponding sequence of ORF234. The total DNA was extracted from the suspected cases of Macrobrachium rosenbergii and used as a template for PCR amplification The specific fragment of about 885bp was cloned into the plasmid vector pET-28a (+) for sequencing and analysis. The results showed that the amplified sequence encoded a total of 294 amino acids with a predicted molecular weight of 34 kDa and 99% identity with the corresponding region of the accession sequence in GenBank. The protein encoded by this sequence has a globular structure in the 22-96 amino acid positions and a conserved region of nearly 130 amino acids homologous to the DUF1335 protein in the 142-272 amino acid positions. Its function is unknown. At the same time, we also carried out sequence analysis and protein prediction on cloned fragments to further study the function of proteins and the prevention and treatment of the virus disease.