Expression and immunogenicity analysis of two iron-regulated outer membrane proteins of Vibrio parahaemolyticus.
Genes of two iron-regulated outer membrane proteins of Vibrio parahaemolyticus zj2003, a pathogenic strain isolated from large yellow croaker (Pseudosciaena crocea), psuA and pvuA, were cloned and expressed as N-terminal His(6)-tagged proteins in Escherichia coli BL(21)(DE(3)). The recombinant fusion proteins were purified with nickel chelate affinity chromatography. To analyze the immunogenicity of the proteins, groups of large yellow croaker were immunized with the purified recombinant psuA, pvuA or both, by intraperitoneal injection. Antibody response was assessed by enzyme-linked immunosorbent assay. Titers to the recombinant proteins increased from log(2) 3.25 to log(2) 9.80, 4-8 weeks following immunization. The relative percent survival of the groups vaccinated with psuA, pvuA, or a combination of the two, reached 50%, 62.5% and 75%, respectively. Western blot analysis was carried out with the serum from unvaccinated survival fish after infection. Both recombinant proteins were detected, indicating that these two proteins of V. parahaemolyticus zj2003 were immunogenic and could produce synergistic effects during in vivo infection, and they might be considered as important components for developing an aquaculture vaccine against this pathogen.
Mao
Zhijuan
Z
College of Biomedical Engineering and Instrument Science, Zhejiang University, Hangzhou 310027, China.
Yu
Lian
L
You
Zhenqiang
Z
Wei
Yongwei
Y
Liu
Yan
Y
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D004926:Escherichia coli; D010473:Perciformes / Q000382:microbiology; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D044149:Vibrio alginolyticus; D014736:Vibrio parahaemolyticus / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D041261:Vibrio vulnificus
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D011956:Receptors, Cell Surface; C083821:iroA protein, bacteria; C449643:pvuA protein, Vibrio