摘要
[目的]了解wsv006蛋白结构变异情况,并进一步为其蛋白功能研究提供参考资料。[方法]以2016和2017年在全国18个地区采集的WSSV阳性样品为模板,采用PCR方法,用特异性引物扩增wsv006基因,经连接转化后进行测序,并对测序结果进行分析。[结果]在2016年的47份样本中有9个样本,包括河北黄骅、山东日照、河北沧州和河北唐山4个地区的样本中出现了wsv006目的条带,检出率为19%。在2017年的39份WSSV阳性样本中有16个样本,包括福建霞美镇、河北黄骅、广东湛江、山东滨州、山东东营和上海6个地区的样本中出现了wsv006的目的条带,检出率为41%。氨基酸序列比对结果显示,扩增的wsv006编码氨基酸变异包括氨基酸替换、插入、缺失和提前终止翻译,其中插入的主要是脯氨酸(P)和苏氨酸(T),插入位点在第151～166位,处于PT的交叉重复区域;终止翻译是由于核苷酸第541位插入了GAGG,在转录过程中形成了终止密码子。[结论]wsv006氨基酸序列存在明显变异。
[Objective]To understand the variations of wsv006,and provide reference for further study of its protein function.[Method]WSSV-positive samples collected in 18 disease outbreak areas during 2016 and 2017 were investigated.The wsv006 gene was amplified by PCR with specific primers and sequenced after ligation and transformation.[Result]The target band of wsv 006 was amplified only in 9 samples from Huanghua,Rizhao,Cangzhou and Tangshan,respectively among the 47 samples collected in 2016,with a detection rate of 19%.For the 39 WSSV-positive samples in 2017,there were only 16 samples in six regions including Xiamei Town,Huanghua,Zhanjiang,Binzhou,Dongying and Shanghai amplified the target band of wsv 006 which reached 41%detection rate.Amino acid sequence alignment showed that the variations of amino acid included substitution,insertion,deletion and termination of translation.One of the main variations was the insertion of proline(P)and threonine(T)in the crion of the two amino acids,between the positions of 151-166.Meanwhile,the insertion of GAGG into the nucleotide resulted in frame-shifts with premature termination by introducing stop codon.[Conclusion]There are obvious variations in the amino acid sequence of wsv006 protein.
作者
蔡苗
刘庆慧
万晓媛
黄倢
CAI Miao;LIU Qing-hui;WAN Xiao-yuan(Laboratory for Marine Fisheries Science and Food Production Processes,Qingdao National Laboratory for Marine Science and Technology,Key Laboratory of Maricultural Organism Disease Control,Ministry of Agriculture,Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao,Shandong 266071;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306)
出处
《安徽农业科学》
CAS
2018年第33期75-79,103,共6页
Journal of Anhui Agricultural Sciences
基金
国家自然科学基金项目(31672679)
国家重点基础研究发展计划项目(2012CB114401)
现代农业产业技术体系项目(CARS-47)
作者简介
蔡苗(1991—),女,河北保定人,硕士研究生,研究方向:分子生物学;通讯作者,刘庆慧,研究员,博士,从事对虾病原生物学研究。
