猪传染性胃肠炎病毒福建株M基因的克隆与生物信息学分析

摘要

In this study, we report the cloning and sequencing of M gene of porcine transmissible gastroenteritis virus Fujian strain (TGEV-FJ), and analyze the homology, signal peptide, glycosylation sites, phosphorylation sites, secondary structure of M protein by using bioinformatics software. The complete M gene in TGEV-FJ strain was successfully amplified which has 789 bp, encoding 262 amino acids and then constructed into the recombinant plasmid pGEM-T-M. Sequential analysis and phylogenetic studies demonstrated that the TGEV M gene has highly conservative and consanguineous between TGEV-FJ strain and HN2002 strain. The prediction results showed that M protein in TGEV-FJ strain had a signal peptide cleavage site (ACG16-17 ER), and 3 potential N-glycosylation sites, 6 serine phosphorylation sites, 1 threonine phosphorylation sites and 5 tyrosine phosphorylation sites. The secondary structure of M protein prediction showed that alpha helix, beta sheet and random coil were 11. 59%, 44.93% and 43.48%, respectively. The results of the present study would lay some basic molecular biological foundation for prokaryotic expression of M gene in TGEV-FJ strain, which would subsequently facilitate further research for molecular diagnosis and gene engineering vaccines development.%对猪传染性胃肠炎病毒(porcine transmissible gastroenteritis virus,TGEV)福建分离株(TGEV-FJ 株)M基因进行克隆、测序,并利用生物信息学技术分析TGEV-FJ株M蛋白基因的同源性、信号肽、糖基化位点、磷酸化位点及其二级结构.结果显示,成功扩增出M基因完整目的片段(789 bp),编码262个氨基酸,构建了M基因的克隆重组质粒pGEM-T-M.序列分析与遗传进化树显示,TGEV M基因属高度保守的基因,TGEV-FJ株与HN2002株亲缘较近.TGEV-FJ株M蛋白具有一个潜在信号肽剪切位点(ACG16-17ER),存在3个潜在的N-糖基化位点,丝氨酸、苏氨酸和酪氨酸磷酸化位点分别有6个、1个和5个；TGEV-FJ株M蛋白二级结构α-螺旋区域占11.59％,β-折叠区域占44.93％,无规则卷曲区域占43.48％.