China Animal Husbandry and Veterinary Medicine ›› 2020, Vol. 47 ›› Issue (3): 686-695.doi: 10.16431/j.cnki.1671-7236.2020.03.005

• Biotechnology • Previous Articles     Next Articles

Antigen Epitope Prediction and Recombinant Protein Design and Verification of ORFV B2L Protein

SUN Zhengnan, ZHANG Huanrong   

  1. College of Life Science and Technology, Southwest Minzu University, Chengdu 610041, China
  • Received:2019-08-27 Online:2020-03-20 Published:2020-03-17

Abstract: To obtain the possible antigenic epitopes of ORFV B2L protein and express the recombinant protein efficiently after truncation.DNAstar,Mega 7.0 and other softwares were used to analyze partial ORFV B2L gene sequences recorded by NCBI.The signal peptide,transmembrane region,cytotoxic T cell epitope,helper T cell epitope,B cell epitope and secondary structure of protein were predicted by different online servers.At the same time,tertiary structure prediction was carried out by referring to multiple templates.Antigenic site analysis and recombinant protein design were carried out by synthesizing major predictive data such as epitope,hydrophilicity,surface accessibility and antigen index,the fusion protein was double His·tag.Several possible high-quality epitopes were obtained,which were 88-93,133-138,172-175,251-254,311-313 and 370-377 amino acids,respectively.A recombinant protein containing major predictive epitopes was designed and expressed by fusing two His·tags to link flexible proteins.The purified and renatured expressed proteins existed in the form of polymers.Western blotting results showed that the purified and renatured recombinant proteins had good reactivity.This study acquired the ORFV B2L antigenic sites and constructed the highly efficient prokaryotic expression vector of recombinant B2L protein,providing the bases for further explore the diagnosis reagent and subunit vaccine.

Key words: ORF virus; B2L gene; protein structure; antigen epitope; recombinant protein

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