传染性造血器官坏死病毒-Sn株基质蛋白基因克隆及生物信息学分析  

Cloning and bioinformatics analysis of matrix protein from IHNV-Sn isolate

作　　者：徐黎明[1] 刘洪柏[1] 卢彤岩[1] 

机构地区：[1]中国水产科学研究院黑龙江水产研究所,黑龙江哈尔滨150070

出　　处：《水产学报》2013年第9期1409-1415,共7页Journal of Fisheries of China

基　　金：国家科技支撑计划(2012BAD25B02);中央级公益性科研院所基本科研业务费专项(2012A0504);黑龙江省冷水性鱼类种质资源及增养殖重点开放实验室(201104)

摘　　要：基质蛋白(matrix protein,M)是传染性造血器官坏死病毒(infectious haematopoieticnecrosis virus,IHNV)主要结构蛋白之一,是病毒感染后造成细胞凋亡的主要作用蛋白。为分析IHNV M蛋白的序列及结构特征,研究利用敏感细胞(EPC)培养传染性造血器官坏死病毒-Sn分离株(IHNV-Sn),根据M蛋白基因开放阅读框(open reading frame,ORF)的序列设计引物,利用RT-PCR的方法克隆得到M蛋白全长ORF,并且构建至表达载体pET27b(+)中,构建出pET27-M重组质粒。生物信息学分析结果显示,M蛋白基因的序列长度为588 bp,编码195个氨基酸残基,推导分子量约为21.88 ku,等电点为9.35;氨基酸序列分析表明,M蛋白富含丝氨酸、苏氨酸以及碱性氨基酸,存在丰富的α-螺旋、β折叠和无规则卷曲;M蛋白不含有信号肽;疏水性大于亲水性;没有跨膜区存在;抗原表位预测显示抗原性良好;结构预测显示,不存在N-糖基化位点,存在7个潜在的O-糖基化位点和15个潜在的磷酸化位点。系统进化树分析显示,IHNV-Sn株与美国分离株同为一簇。Matrix protein is one of the structure proteins of infectious haematopoietic necrosis virus（IHNV）,and its synthesis leads to apoptosis or programmed cell death in the transfected cells.To study its structure and function,the open reading frame of matrix protein gene was amplified by RT-PCR from IHNV-Sn isolate and cloned into plasmid pET27b（＋）vector.The structure and characteristics of the gene were studied by means of bioinformatics software.The results are described as follows：the length of the M gene,encoding 195 aa,was 588 bp.The molecular weight was 21.88 ku and the isoelectric point was 9.35.The matrix protein was rich in serine,threonine and alkaline amino acid,and was composed of plenty of α-helix,extended β and coil.The protein is hydrophobic with no signal peptide and transmembrane regions.The antigenicity of matrix protein was good.According to the protein structure prediction,there might be no N-glycosylation sites,and there were 7 O-glycosylation sites and 15 phosphorylation sites in matrix protein.Phylogenetic analysis showed that the IHNV-Sn isolate was in the same branch with the isolates from America.The study established the foundation for genetic background information,pathogenesis,molecular epidemiology research of IHNV.

关 键 词：传染性造血器官坏死病毒-Sn株 基质蛋白 生物信息学分析 

分 类 号：Q939.4[生物学—微生物学] S943[农业科学—水产养殖]