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

doi:10.11843/j.issn.0366-6964.2018.06.017

Abstract

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 foundation for the development of the serodiagnostic methods and immunology research for PEDV.

CLC Number:

WANG Long-bai, WANG Chen-yan, WU Xue-min, CHEN Qiu-yong, CHE Yong-liang, CHEN Ru-jing, ZHOU Lun-jiang. Prokaryotic Expression and Immunogenic Analysis of the Combined M and N Fusion Genes for Variant Porcine Epidemic Diarrhea Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1249-1255.

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Prokaryotic Expression and Immunogenic Analysis of the Combined M and N Fusion Genes for Variant Porcine Epidemic Diarrhea Virus

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