中国兽医科学2018，Vol.48Issue(5)：605-615,11.DOI:10.16656/j.issn.1673-4696.2018.0000
番鸭呼肠孤病毒YB株μB蛋白的原核表达与生物信息学分析
Prokaryotic expression and bioinformatic analysis of μB gene of Muscovy duck reovirus YB strain
摘要
Abstract
To research the homology phylogenetic relationship between μB gene of Muscovy duck reovirus YB strain(MDRV-YB)and the other reference strains,to predict the structure and function of protein μB and to prokaryoticly express MDRV-YB μB gene, in this experiment the μB gene was amplified from virus sample through RT-PCR. Then the recombinant plasmid pET-YB- μB was constructed, the whole CDS sequence of MDRV-YB μB gene was acquired, the biological information of the μB gene was analysed, and its structure and function was predicted by using DNAStar software. The recombinant plasmid pET-YB-μB was transformed into E. coli BL21 (DE3)competent cells for protein expression induced by IPTG. In result,the length of whole CDS sequence of μB gene was 2 031 bp in size, encoding 676 amino acids,and the molecular formula of protein μB was C3217H5105N879O1014S27, without any signal peptides, possibly having 4 N-glycosylation sites and 85 phosphorylation sites. The secondary structure of the protein contained 27.6％ a-helix structure,53.3％ β-fold structure, 10％ turn structure and 9.1％ random coil. Homology analysis showed that μB gene of MDRV-YB strain had a high homology about 99.8％ with MDRV ZJ2000M strain while low homology about 67.3％-69.6％ with other polyclonal strains. The phylog-enetic tree showed that YB strain and MW9710 strain were at the nearest branch. SDS-PAGE and Western-blot analysis results showed that pET-YB- μB could express about 99. lku μB fusion protein in molecular weight,and the protein had great reactinogenicity. This study results laid the foundation for the genetic evolution of MDRV and the protein function research of MDRV μB protein, and also provided the data support for further research to the pathogenic mechanism of MDRV.关键词
番鸭呼肠孤病毒/μB基因/原核表达/生物信息学分析Key words
Muscovy duck reovirus(MDRV)/μBgene/prokaryotic expression/bioinformatic analysis分类
农业科技引用本文复制引用
陈强,吴异健,张英扬,蔡栋灵,刘珍妮,骆钰,林绍青,李鸿文,周五朵,朱二鹏..番鸭呼肠孤病毒YB株μB蛋白的原核表达与生物信息学分析[J].中国兽医科学,2018,48(5):605-615,11.基金项目
福建省农业科技重大专项(2014NZ0002-4):白羽肉鸡主要疾病防控关键技术研究及应用 （2014NZ0002-4）
福建农林大学科技创新专项基金(CXZX2016016) （CXZX2016016）
猴头菇多糖对MDRV感染雏番鸭肠黏膜淋巴细胞归巢过程的影响及其机制研究 （）
