title: Evaluation of optimal culture conditions for recombinant ghost bacteria vaccine production with the antigen of Streptococcus iniae GAPDH.
abstract: For the production of ghost bacteria vaccine to prevent the streptococcal disease in aquaculture fish species, a double cassettes vector was constructed and cloned in Escherichia coli DH5alpha. Ghost bacteria vaccine production from Escherichia coli DH5alpha/pHCE-InaN-GAPDH-Ghost 37 SDM (SIG) was maximized at a glucose concentration of 1 g/l, agitation of 300 rpm, and aeration of 1 vvm. The maximal efficiency of ghost bacteria formation was obtained at the mid-exponential phase (OD600=2.0) with the concentration of 0.77 g/l for SIG. The molecular mass of GAPDH was detected at 67 kDa with the insoluble fraction, by SDS-PAGE and Western blot. The protective efficacy of ghost bacteria vaccine was evaluated by challenge test using olive flounder. The cumulative mortalities of the positive control, formalin-killed cell (FKC) vaccine, and SIG vaccine immunized groups were 91% , 74% , and 57% , respectively. These results suggest that SIG vaccine showed efficacy as a vaccine and had a higher potential to induce protective antibodies than did FKC vaccine.
authors.affiliation: Department of Biotechnology, Pukyong National University, Busan 608-737, Korea.
authors.forename: Chae-Hun
authors.initials: CH
authors.lastname: Ra
authors.forename: Yeong-Jin
authors.initials: YJ
authors.lastname: Kim
authors.forename: So-Jin
authors.initials: SJ
authors.lastname: Park
authors.forename: Chang-Wha
authors.initials: CW
authors.lastname: Jeong
authors.forename: Yoon-Kwon
authors.initials: YK
authors.lastname: Nam
authors.forename: Ki-Hong
authors.initials: KH
authors.lastname: Kim
authors.forename: Sung-Koo
authors.initials: SK
authors.lastname: Kim
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005393:Fish Diseases / Q000276:immunology / Q000382:microbiology*; D005399:Fishes; D013290:Streptococcal Infections / Q000276:immunology / Q000662:veterinary*; D013291:Streptococcus / Q000276:immunology*
chemical_list: D000942:Antigens, Bacterial; D001428:Bacterial Vaccines