畜牧兽医学报2018，Vol.49Issue(6)：1249-1255,7.DOI:10.11843/j.issn.0366-6964.2018.06.017
变异猪流行性腹泻病毒M和N融合双基因的原核表达及表达产物免疫原性分析
Prokaryotic Expression and Immunogenic Analysis of the Combined M and N Fusion Genes for Variant Porcine Epidemic Diarrhea Virus
摘要
Abstract
The aim of this study was to construct a double gene fusion plasmid of the membrane(M)and nucleocapsid(N)genes of variant porcine epidemic diarrhea virus(PEDV)and analyze the immunogenicity of the expressed proteins.The M and N genes were amplified by PCR from PEDV variant isolate,FJFQ2014(GenBank No.KJ646580).First,the M gene was cloned into pEASY-Blunt E2(pE2)vector to construct pE2-M plasmid.Then the amplified N gene and E2-M gene of expression vector pE2-M were cloned into the prokaryotic expression vector,resulting a recombinant plasmid pE2-M-N.The plasmid was transformed to Transetta(DE3)for protein expression.The expressed product was identified by SDS-PAGE and Western blot.The BLAB/c mice were immunized with the purified PEDV M-N protein and the antibody titer was determined by ELISA.The results indicated that PEDV M-N fusion protein was successfully expressed in Transetta(DE3)and had immunogenicity,and induced anti-PEDV antibody in mice.The pE2-M-N plasmid of variant PEDV was constructed and expressed successfully.This study lays a foun-dation for the development of the serodiagnostic methods and immunology research for PEDV.关键词
猪流行性腹泻病毒/PEDV/M基因/N基因/双基因表达Key words
porcine epidemic diarrhea virus/PEDV/M gene/N gene/expression of two genes分类
农业科技引用本文复制引用
王隆柏,王晨燕,吴学敏,陈秋勇,车勇良,陈如敬,周伦江..变异猪流行性腹泻病毒M和N融合双基因的原核表达及表达产物免疫原性分析[J].畜牧兽医学报,2018,49(6):1249-1255,7.基金项目
公益类科研院所专项(2018R1023-4) （2018R1023-4）
福建省自然科学基金项目(2015J01112) （2015J01112）
公益类科研院所专项(2017R1023-8) （2017R1023-8）
