Recombinant VP1 protein of duck hepatitis virus 1 expressed in Pichia pastoris and its immunogenicity in ducks

被引:6
作者
Wang, C. [1 ,2 ]
Li, X. K. [1 ]
Wu, T. C. [1 ]
Wang, Y. [1 ]
Zhang, C. J. [1 ]
Cheng, X. C. [1 ]
Chen, P. Y. [2 ]
机构
[1] Henan Univ Sci & Technol, Coll Anim Sci & Technol, Luoyang 471003, Henan Province, Peoples R China
[2] Nanjing Agr Univ, Chinas Dept Agr, Key Lab Anim Dis Diag & Immunol, Nanjing 210095, Jiangsu, Peoples R China
基金
中国国家自然科学基金;
关键词
duck hepatitis virus 1; genotype C; VP1; protein; Pichia pastoris; duck; immunogenicity; MOUTH-DISEASE VIRUS; MOLECULAR ANALYSIS; ANTIGENIC VARIATION; HUMAN PARECHOVIRUS; IMMUNE-RESPONSE; IN-VITRO; SEROTYPE; TYPE-1; GENUS; PEPTIDE;
D O I
10.4149/av_2014_04_333
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The VP1 gene of duck hepatitis virus type 1 (DHV-1) strain VJ09 was amplified by reverse transcription PCR from the liver of a duckling with clinical symptoms of viral hepatitis. The resulting VP1 cDNA was 720 bp in length and encoded a 240-amino-acid protein. In VP1 gene-based phylogenetic analysis, the VJ09 strain grouped with DHV-1 genotype C. The VP1 gene was inserted into the expression vector pPICZ alpha A and expressed in Pichia pastoris. The expressed VP1 protein was purified and identified by western blot analysis. To evaluate the recombinant VP1's immunogenic potential in ducklings, the antibodies raised in the immunized ducklings were titrated by ELISA, and lymphocyte proliferation and virus neutralization assays were performed. The results show that the recombinant VP1 protein induced a significant immune response in ducklings and this could be a candidate for the development of a subunit vaccine against DHV-1 genotype C.
引用
收藏
页码:333 / 339
页数:7
相关论文
共 35 条
[1]   PROTECTION AGAINST FOOT-AND-MOUTH-DISEASE BY IMMUNIZATION WITH A CHEMICALLY SYNTHESIZED PEPTIDE PREDICTED FROM THE VIRAL NUCLEOTIDE-SEQUENCE [J].
BITTLE, JL ;
HOUGHTEN, RA ;
ALEXANDER, H ;
SHINNICK, TM ;
SUTCLIFFE, JG ;
LERNER, RA ;
ROWLANDS, DJ ;
BROWN, F .
NATURE, 1982, 298 (5869) :30-33
[2]   Arginine-glycine-aspartic acid motif is critical for human parechovirus 1 entry [J].
Boonyakiat, Y ;
Hughes, PJ ;
Ghazi, F ;
Stanway, G .
JOURNAL OF VIROLOGY, 2001, 75 (20) :10000-10004
[3]   Heterologous protein expression in the methylotrophic yeast Pichia pastoris [J].
Cereghino, JL ;
Cregg, JM .
FEMS MICROBIOLOGY REVIEWS, 2000, 24 (01) :45-66
[4]  
COLLEN T, 1991, J IMMUNOL, V146, P749
[5]   Recombinant protein expression in Pichia pastoris [J].
Cregg, JM ;
Cereghino, JL ;
Shi, JY ;
Higgins, DR .
MOLECULAR BIOTECHNOLOGY, 2000, 16 (01) :23-52
[6]   Molecular analysis of duck hepatitis virus type 1 [J].
Ding, Chunyu ;
Zhang, Dabing .
VIROLOGY, 2007, 361 (01) :9-17
[7]   Structure of Foot-and-mouth disease virus serotype A1061 alone and complexed with oligosaccharide receptor:: receptor conservation in the face of antigenic variation [J].
Fry, EE ;
Newman, JWI ;
Curry, S ;
Najjam, S ;
Jackson, T ;
Blakemore, W ;
Lea, SM ;
Miller, L ;
Burman, A ;
King, AMQ ;
Stuart, DI .
JOURNAL OF GENERAL VIROLOGY, 2005, 86 :1909-1920
[8]   INVITRO ISOLATION, PROPAGATION, AND CHARACTERIZATION OF DUCK HEPATITIS-VIRUS TYPE-III [J].
HAIDER, SA ;
CALNEK, BW .
AVIAN DISEASES, 1979, 23 (03) :715-729
[9]   Codon optimization, expression, and characterization of an internalizing anti-ErbB2 single-chain antibody in Pichia pastoris [J].
Hu, Siyi ;
Li, Liangwei ;
Qiao, Jingjuan ;
Guo, Yujie ;
Cheng, Liansheng ;
Liu, Jing .
PROTEIN EXPRESSION AND PURIFICATION, 2006, 47 (01) :249-257
[10]   Isolation and identification of a novel human parechovirus [J].
Ito, M ;
Yamashita, T ;
Tsuzuki, H ;
Takeda, N ;
Sakae, K .
JOURNAL OF GENERAL VIROLOGY, 2004, 85 :391-398