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Characterization of OmpK, GAPDH and their fusion OmpK–GAPDH derived from Vibrio harveyi outer membrane proteins: their immunoprotective ability against vibriosis in large yellow croaker ( Pseudosciaena crocea )
Author(s) -
Zhang C.,
Yu L.,
Qian R.
Publication year - 2007
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2007.03386.x
Subject(s) - glyceraldehyde 3 phosphate dehydrogenase , biology , recombinant dna , microbiology and biotechnology , vibrio harveyi , fusion protein , vibrio infections , bacterial outer membrane , escherichia coli , vibrionaceae , bacteria , biochemistry , dehydrogenase , vibrio , enzyme , gene , genetics
Aim: To investigate the immunoprotection of three recombinant proteins derived from the Vibrio harveyi outer membrane proteins (OMPs) OmpK, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) and their fusion OmpK–GAPDH as vaccine candidates from vibriosis of large yellow croaker ( Pseudosciaena crocea ). Methods: The ompK gene, of which the leader sequence was omitted, was fused with the gapdh gene. Three recombinant proteins r‐OmpK, r‐GAPDH and r‐OmpK–GAPDH were expressed and purified. Western blots were carried out to detect the specificity of the antibodies raised against the recombinant proteins; Fish were immunized with recombinant proteins and challenged by native V. harveyi . The immunoresponse to the recombinant proteins were determined by ELISA and phagocytic activity assay. Conclusions: The fusion protein r‐OmpK–GAPDH can afford greater protection against the wild V. harveyi than r‐OmpK or r‐GAPDH alone or their mixture in humoral and cellular immunity , indicating that OmpK and GAPDH could produce a synergistic immunoprotection against vibriosis of large yellow croaker ( Pseudosciaena crocea ) when fused into OmpK–GAPDH with a linker. Significance and Impact of the Study: It has been realized that a multi‐component OMP antigen can induce a higher frequency of immune effectors than a single OMP. The results presented here bring forth a good suggestion for the subunit vaccine design based on the OMPs of gram‐negative pathogens.