以WWSV ORF234片段诊断罗氏沼虾白斑病毒病及片段分析  

Diagnosis of White Spot Syndrome Virus Disease by ORF234 of Macrobrachium rosenbergii and Sequence Analysis

作　　者：杨贺[1] 王晓兰[2] 陈辉 韩凤丽[1] 贡成良[1] 曹广力[1] 薛仁宇[1] 

机构地区：[1]苏州大学生命科学学院,江苏苏州215123 [2]常熟理工学院,江苏常熟215500 [3]江苏省水生动物疫病控制中心,江苏南京210036

出　　处：《水生态学杂志》2008年第6期101-106,共6页Journal of Hydroecology

基　　金：苏州市攻关课题(编号:SNZ0518)

摘　　要：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。A pair of primers was designed based on white spot syndrome virus （WSSV） released from GenBank to amplify a fragment with total DNA template from Macrobrachium rosenbergii which was suspicious of being infected with WSSV. The fragment then was cloned into vector of pet -28a（ ＋ ）. Nucleotide sequence analysis showed the sequence amplified was 99% identity with the ORF234 of total sequence of WSSV from GeneBank, It is believed that the shrimp was infected with WSSV. The sequence encoded 294 amino acids, the estimated molecular weight was 34KDa. Protein encoded by the sequence has a globular structure at 22 -96 amino acid, and at 142 -272 amino acid there was a nearly 130 amino aeid residues conservative district homologous with DUF1335 ,whieh funetion was unknown. Meanwhile sequenee analysis and protein predietion has been studied for further reseaeh of the protein function and the control of virus disease.

关 键 词：对虾白斑综合症病毒 ORF234基因 克隆 序列分析 罗氏沼虾 

分 类 号：Q74[生物学—分子生物学]