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Vibrio harveyi is an important marine pathogen that can infect a number of aquaculture species.V. harveyi degQ (degQVh ), the gene encoding a DegQ homologue, was cloned from T4, a pathogenicV. harveyi strain isolated from diseased fish. DegQVh was closely related to the HtrA family members identified in otherVibrio species and could complement the temperature-sensitive phenotype of anEscherichia coli strain defective indegP . Expression ofdegQVh in T4 was modulated by temperature, possibly through the σE -like factor. Enzymatic analyses demonstrated that the recombinant DegQVh protein expressed in and purified fromE. coli was an active serine protease whose activity required the integrity of the catalytic site and the PDZ domains. The optimal temperature and pH of the recombinant DegQVh protein were 50°C and pH 8.0. A vaccination study indicated that the purified recombinant DegQVh was a protective immunogen that could confer protection upon fish against infection byV. harveyi . In order to improve the efficiency of DegQVh as a vaccine, a genetic construct in the form of the plasmid pAQ1 was built, in which the DNA encoding the processed DegQVh protein was fused with the DNA encoding the secretion region of AgaV, an extracellular β-agarase. TheE. coli strain harboring pAQ1 could express and secrete the chimeric DegQVh protein into the culture supernatant. Vaccination of fish with viableE. coli expressing chimericdegQVh significantly (P &lt; 0.001) enhanced the survival of fish againstV. harveyi challenge, which was possibly due to the relatively prolonged exposure of the immune system to the recombinant antigen produced constitutively, albeit at a gradually decreasing level, by the carrier strain.

Characterization of DegQ Vh , a Serine Protease and a Protective Immunogen from a Pathogenic Vibrio harveyi Strain

Characterization of DegQ _Vh , a Serine Protease and a Protective Immunogen from a Pathogenic Vibrio harveyi Strain