中国畜牧兽医2017，Vol.44Issue(12)：3585-3591,7.DOI:10.16431/j.cnki.1671-7236.2017.12.027
传染性法氏囊病病毒C4毒株VP2基因的原核表达及生物信息学分析
Prokaryotic Expression and Bioinformatics Analysis of VP2 Gene of Infectious Bursal Disease Virus C4 Strain
摘要
Abstract
This study was aimed to investigate the relationship between the virulence characteristics of infectious bursal disease virus(IBDV) C4 strain and its VP2 amino acid sequence.The RNA of IBDV C4 strain was extracted,and its VP2 gene was amplified by RT-PCR.VP2 nucleotide sequences and deduced amino acids of different virulent IBDV strains were compared.At the same time,prokaryotic expression vector pET-32a(+) was used to express the VP2 gene.The expression of recombinant VP2 protein was detected by SDS-PAGE and Western blotting.The results showed that the VP2 gene of IBDV C4 strain belonged to the very virulent infectious bursal disease virus (vvIBDV) in evolutionary relationship,the VP2 nucleotides homology between IBDV C4 strain and other vvIBDV strains were 98.1％ to 98.7％,and there were no mutations in S-W-S-A-SGS (326-332 amino acids) and 222(A),256(I),294(I) and 299(S).The VP2 amino acid sequence of IBDV C4 strain was consistent with the characteristics of other vvIBDV strains.However,there were three differences amino acids sites at 201(D/G),281(G/R) and 313(V/A) between the amino acids of the C4 strain and the very virulent strain UK661.And the change of 281(R) was in the small hydrophilic region of 279 to 290,which was related to the antigenicity of the virus;The recombinant VP2 protein molecular weight expressed in Escherichia coli BL21 was about 67 ku.This study provided a basis for further research on antigenic changes resulting from amino acid variation of 201(G),281 (R) and 313(A).These results indicated that the VP2 gene of the IBDV C4 strain was consistent with the major characteristics of the vvIBDV strain VP2 gene.The difference of three amino acid sites in the vvIBDV strain C4 might be related to the evolution of virulence of IBDV strain in China.关键词
传染性法氏囊病病毒(IBDV)/VP2/分子特征/原核表达Key words
infectious bursal disease virus (IBDV)/VP2/molecular characterization/prokaryotic expression分类
农业科技引用本文复制引用
王洁琼,赵玉杰,周云飞,黄宗梅,陈盼盼,周薇帆,刘琳,李新生..传染性法氏囊病病毒C4毒株VP2基因的原核表达及生物信息学分析[J].中国畜牧兽医,2017,44(12):3585-3591,7.基金项目
国家十三五科技专项(2017YFD0500701) （2017YFD0500701）
