Prokaryotic expression and purification of PEDV N protein and analysis of B cell antigen epitope

doi:10.3969/j.issn.1004-1524.2020.05.02

›› 2020, Vol. 32 ›› Issue (5): 762-769.DOI: 10.3969/j.issn.1004-1524.2020.05.02

• Animal Science • Previous Articles Next Articles

Prokaryotic expression and purification of PEDV N protein and analysis of B cell antigen epitope

GUO Fucheng^1,2, JIN Li^1,2, SU Qiang^1,2, SU Yuxin^1,2, LI Fanglin^1,2, CHEN Shien^1,2, MA Xiaoxia^1,2,*

1. Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China;
2. College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China

Received:2019-10-08 Online:2020-05-25 Published:2020-05-29

Abstract

Abstract: The aim of this study was to clone and construct the porcine epidemic diarrhea virus (PEDV) N gene and to predict the epitope properties of its B cells. The N gene of PEDV was amplified by PCR and cloned into the prokaryotic expression vector pET-28a(+).The positive clone was obtained by enzyme digestion, and the positive clone was expressed in the expression strain E. coli BL21 (DE3). High purity expressed proteins were obtained by optimizing expression conditions and purification conditions. The 3D structure of PEDV-N protein was established by bioinformatics homology modeling software Swiss-Pdb Viewer and the secondary structure, antigenicity, hydrophilicity and surface probability of PEDV-N protein were predicted according to bioinformatics software DNA-Star, and the antigen epitopes of its B cells were predicted by comprehensive analysis. It is predicted that there were 13 B cell dominant epitopes in the amino acid sequence of PEDV-N protein. The research results will further provide a theoretical basis for the application of PEDV-N protein expression products in vitro and the development of genetically engineered vaccines.

Key words: porcine epidemic diarrhea virus, N protein, purification, secondary structure, B-cell epitope prediction

CLC Number:

S825.28

GUO Fucheng, JIN Li, SU Qiang, SU Yuxin, LI Fanglin, CHEN Shien, MA Xiaoxia. Prokaryotic expression and purification of PEDV N protein and analysis of B cell antigen epitope[J]. , 2020, 32(5): 762-769.

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Prokaryotic expression and purification of PEDV N protein and analysis of B cell antigen epitope

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