CDV强毒BJ16B2株H基因在昆虫杆状病毒中的表达与鉴定

doi:10.16431/j.cnki.1671-7236.2018.08.001

Abstract

Abstract:

To obtain the native conformation and antigenicity of hemagglutinin (H) protein of canine distemper virus (CDV) virulent strain,the H gene which referred to BJ16B2 strain was accurately amplified by RT-PCR and cloned into pFastBac^TMHTA vector.The recombinant plasmid was confirmed by restriction enzyme digestion and DNA sequencing,H gene was analyzed and the recombinant plasmid was transformed into E.coli DH10Bac^TM competent cell,and constructed shuttle vector,which was transfected into Sf9 cell to save baculovirus expressing H protein.To determine the antigenicity of recombinant protein,Western blotting and IFA analysis were used to detect the interesting protein.The results demonstrated that H gene belongs to Asia-1 virulent strain,and the homology of nucleotide and amino acid among the reference strains was 89.8% to 99.3% and 89.6%to 99.5%,respectively.The fusion protein generated by recombinant baculovirus was reacted with both His-tag monoclonal antibodies and CDV positive serum by Western blotting test,and presented specific green fluorescence in IFA test (68 ku),which showed that this protein had sufficient antigenicity.The resluts served as a strong foundation for the the subsequent analysis of protein structure and antigenicity differences,the preparation of monoclonal antibodies and the development of CDV subunit vaccine.

Key words: canine distemper virus (CDV); H gene; insect baculovirus expression system

CLC Number:

S855.3

CHEN Meirong, LIN Weidong, SONG Tianqi, XIN Ting, GUO Xiaoyu, HUANG Kehe, JIA Hong. Expression and Identification of H Gene of CDV BJ16B2 Strain in Insect Baculovirus Expression System[J]. , 2018, 45(8): 2041-2048.

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Expression and Identification of H Gene of CDV BJ16B2 Strain in Insect Baculovirus Expression System

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