Immunological Identification and Characterization of Extracellular Serine Protease-Like Protein Encoded in a Putative espP2 Gene of Haemophilus parasuis
文献类型: 外文期刊
第一作者: Zhang, Nian-Zhang
作者: Zhang, Nian-Zhang;Chu, Yue-Feng;Gao, Peng-Cheng;Zhao, Ping;He, Ying;Lu, Zhong-Xin
作者机构:
关键词: cloning;ESP-like protein;expression;Haemophilia parasuis;immunogenicity
期刊名称:JOURNAL OF VETERINARY MEDICAL SCIENCE ( 影响因子:1.267; 五年影响因子:1.348 )
ISSN: 0916-7250
年卷期: 2012 年 74 卷 8 期
页码:
收录情况: SCI
摘要: Haemophilus parasuis is known to produce a group of virulence-associated autotransporter (AT) proteins, VtaAs; however, no other ATs have been characterized yet. On the basis of the reported sequence of a putative espP2 gene for extracellular serine protease (ESP)-like protein of H. parasuis, this putative AT gene was successfully amplified from H. parasols serotype 5 field strain HPS0819, cloned and sequenced. The confirmed ORF sequence showed 100% identity with the reported putative espP2 gene. The recombinant ESP-like protein purified from Escherichia coli with a pET expression system was used for immunological characterization. An approximately 85 kDa antigen was detected in cultured H. parasuis by using antiserum to the purified ESP-like protein, and antibodies against the recombinant ESP-like protein were detected in a selected serum from pigs with experimental H. parasuis infection. The results indicated that H. parasols could produce ESP-like protein in vitro and in vivo. In an immune protection study using guinea pigs, 6 out of 10 animals immunized with the recombinant ESP-like protein survived after challenge with 5 x 10(9) bacteria of strain HPS0819, whereas 7 out of 10 animals immunized with formalin-inactivated H0819 bacterin survived after challenge. The results suggest that ESP-like protein could be one of the vaccine antigen candidates for H. parasuis infection.
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