猪源戊型肝炎病毒ORF2蛋白生物信息学分析及真核表达研究

doi:10.16431/j.cnki.1671-7236.2023.05.023

Abstract

Abstract: 【Objective】 The purpose of this study was to analyze the structure and function of ORF2 protein of Swine hepatitis E virus (sHEV) genotype 4,screen the gene fragment with protective antigen and express,so as to provide a candidate protein for the study of its potential protective antigen.【Method】 The bioinformatics analysis of sHEV genotype 4 ORF2 protein was carried out.The gene fragment ORF2 (128/140) with potential protective antigen protein was selected,amplified by PCR and digested,which cloned into insect cell expression vector,and transfected into sf9 cells.The expressed protein p128/p140 were analyzed by Western blotting.【Result】 Bioinformatics analysis showed that the ORF2 protein of sHEV genotype 4 had two stable proteins (p128 and p140),which contained 496 and 476 amino acids,respectively.Both of them were stable proteins,without transmembrane regions,and the secondary structure of the protein was mainly random coil.p128 protein had 11 T cell epitopes and 21 B cell epitopes,and p140 protein had 14 T cell epitopes and 18 B cell epitopes.Insect cell expression vector of ORF2(128) and ORF2(140) genes were successfully constructed,and transfected into sf9 cells,Western blotting results showed that the expressed proteins could be recognized by His-tag monoclonal antibody and positive serum of sHEV antibody.【Conclusion】 p128 and p140 proteins with potential protective antigen of sHEV were found in this study,and the two proteins were expressed in sf9 cells with immune activity,which provide materials for further study of the function of sHEV ORF2 protein and the development of new vaccines.

Key words: Swine hepatitis E virus (sHEV); ORF2 protein; bioinformatics analysis; insect cell; expression

CLC Number:

ZHUO Na, ZHU Zhongwu, LI Jing, HU Shixiang, WANG Jianchang, LIN Hua, CHEN Chaolin, HAN Diangang, AI Jun, CHEN Peifu. Bioinformatics Analysis and Eukaryotic Expression of ORF2 Protein of Swine Hepatitis E Virus[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1959-1970.

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Bioinformatics Analysis and Eukaryotic Expression of ORF2 Protein of Swine Hepatitis E Virus

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