急性病毒性坏死病毒魁蚶株IAP-86基因全长cDNA克隆和生物信息学分析  

Full Length cDNA Cloning and Bioinformatics Analysis of Acute Viral Necrosis Virus IAP-86 Gene From Anadara uropygimelana

作　　者：张帅[1] 王崇明[1] 岳志芹[2] 宋晓玲[1] 白昌明[1] 尹伟力[2] 黄倢[1] 

机构地区：[1]农业部海洋渔业可持续发展重点实验室,中国水产科学研究院黄海水产研究所,青岛266071 [2]山东出入境检验检疫局检验检疫技术中心,青岛266002

出　　处：《渔业科学进展》2015年第1期85-90,共6页Progress in Fishery Sciences

基　　金：现代农业产业技术体系建设专项(CARS-48);国家质检总局科研项目(2013IK040)共同资助

摘　　要：为深入探讨急性病毒性坏死病毒(Acute Viral Necrosis Virus,AVNV)魁蚶株的致病机理以及IAP-86蛋白的功能,从感染AVNV的濒死魁蚶外套膜中提取总RNA,反转录获得c DNA。根据NCBI公布的AVNV全基因组序列中ORF86序列设计两对反向套式引物,通过c DNA末端快速扩增(RACE)技术获得ORF86 5?端和3?端的非编码区,拼接获得全长c DNA序列。Blast序列比对显示,该基因与牡蛎疱疹病毒的同源性为100%,与栉孔扇贝AVNV的同源性为99%,并存在重叠基因。生物信息学分析预测,该蛋白不含信号肽,不存在跨膜区,最大疏水指数为1.800,最小疏水指数为–3.456。该蛋白存在8个潜在的磷酸化位点(包括5个丝氨酸、1个苏氨酸和2个酪氨酸),存在1个潜在的O-糖基化位点,不存在潜在的N-糖基化位点;其抗原表位主要集中在8-11、14-16、28-39、75-76、88-95、97-100和147-158位氨基酸。推测该株病毒可能为牡蛎疱疹病毒的变异株,并且基因重叠在该类病毒进化过程中可能扮演重要角色。Acute viral necrosis virus （AVNV） was reported as the causative agent for summer mass mortality of adult Zhikong scallop （Chlamys farreri） that has been widely cultured along northern China coast. To understand the pathogenesis and the function of IAP-86, a strain of acute viral necrosis virus （AVNV） was isolated from Anadara uropygimelana. RNA was extracted from the mantle of moribund A. uropygimelana that was infected with AVNV, and cDNA ＇was obtained by reverse transcription. Two pairs of nested reverse primer were designed according to the ORF86 sequence of AVNV complete genome sequence that registered in NCBI. The non-coding region of 5＇ and 3＆#39; end of the ORF86 were amplified using the designed primers by rapid amplification of cDNA ends （RACE） technique, and the full-length cDNA sequences were spliced. Blast sequence alignment illustrated that this gene has 100% homology with oyster herpetovirus and 99% with the AVNV. Moreover, overlapping genes were found in the cDNA sequence. Bioinformatics analysis indicated that the protein contains neither a signal peptide nor a transmembrane region. The maximum hydrophobic index was 1.800 and the minimum hydrophobic index was -3.456. There are eight potential phosphorylation sites （including five serine sites, two tyrosine sites and one threonine site）, a potential O-glycosylation site, but no potential N-glycosylation site. The epitope were mainly located on the amino acids of 8-11, 14-16, 28-39, 75-76, 88-95, 97-100 and 147-158. Results suggest that the virus may be a strain of oyster herpetovirus, and gene overlapping may play an important role in the virus evolution.

关 键 词：急性病毒性坏死病毒 魁蚶 细胞凋亡抑制蛋白 c DNA末端快速扩增技术 生物信息学 

分 类 号：S944[农业科学—水产养殖]