title: Cloning, expression, and immunogenicity of Flavobacterium columnare heat shock protein dnaJ.
abstract: The Flavobacterium columnare heat shock protein (HSP) gene dnaJ* was isolated, cloned, expressed, and used as an antigen in a recombinant vaccine strategy for channel catfish Ictalurus punctatus. The F. columnare dnaJ* sequence was obtained from genomovars I and II and showed intraspecies variability. Recombinant protein was expressed and purified from Escherichia coli cultures and injected intraperitoneally (12 microg of purified DnaJ/fish) into fingerling channel catfish. In addition, induced (expressing the recombinant DnaJ) and uninduced (no recombinant protein being produced) E. coli cultures were also used to immunize fish. At 28 d postimmunization, antibody response was evaluated and the fish were challenged with F. columnare. A specific immune response against DnaJ was observed in fish immunized with DnaJ or E. coli cultures expressing DnaJ. No protection against the disease, however, was observed in F. columnare-challenged fish that had been immunized with DnaJ. Some level of protection was observed in fish immunized with uninduced and induced E. coli lysates. Although HSPs have been shown to be immunodominant and good candidates for subunit vaccines in other animals, DnaJ failed to protect against columnaris disease in channel catfish.
authors.affiliation: Department of Fisheries and Allied Aquacultures, Auburn University, 217 Swingle Hall, Auburn, Alabama 36849, USA.
authors.forename: Oscar
authors.initials: O
authors.lastname: Olivares-Fuster
authors.forename: Jeffery S
authors.initials: JS
authors.lastname: Terhune
authors.forename: Craig A
authors.initials: CA
authors.lastname: Shoemaker
authors.forename: Covadonga R
authors.initials: CR
authors.lastname: Arias
mesh_terms: D000818:Animals; D001428:Bacterial Vaccines; D003001:Cloning, Molecular; D005393:Fish Diseases / Q000276:immunology / Q000382:microbiology; D045826:Flavobacteriaceae Infections / Q000382:microbiology / Q000517:prevention & control / Q000662:veterinary; D005417:Flavobacterium / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D050956:HSP40 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007059:Ictaluridae; D010802:Phylogeny; D011994:Recombinant Proteins
chemical_list: D001428:Bacterial Vaccines; D050956:HSP40 Heat-Shock Proteins; D011994:Recombinant Proteins