Evaluation of optimal culture conditions for recombinant ghost bacteria vaccine production with the antigen of Streptococcus iniae GAPDH.
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.
Ra
Chae-Hun
CH
Department of Biotechnology, Pukyong National University, Busan 608-737, Korea.
Kim
Yeong-Jin
YJ
Park
So-Jin
SJ
Jeong
Chang-Wha
CW
Nam
Yoon-Kwon
YK
Kim
Ki-Hong
KH
Kim
Sung-Koo
SK
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*
D000942:Antigens, Bacterial; D001428:Bacterial Vaccines