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http://dx.doi.org/10.5352/JLS.2010.20.12.1743

Roles of Glyceraldehyde-3-Phosphate Dehydrogenase in Edwardsiella tarda Pathogenesis  

Yu, Jong-Earn (Department of Microbiology, Pusan National University)
Oh, Young-Eun (Department of Microbiology, Pusan National University)
Lee, Tae-Ho (Department of Microbiology, Pusan National University)
Kang, Ho-Young (Department of Microbiology, Pusan National University)
Publication Information
Journal of Life Science / v.20, no.12, 2010 , pp. 1743-1749 More about this Journal
Abstract
A research group demonstrated that the 37 kDA protein of Edwardsiella tarda, a causing causative agent of edwardsiellosis in fish, exhibited high antigenicity in Japanese flounder. The research group also showed that the N-terminus amino acid sequences of the 37 kDa protein were mapped to the N-terminus of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Using degenerated primer sets based on the known N-terminus sequence, the corresponding E. tarda DNA was amplified and cloned. The nucleotide sequences of the cloned gene revealed high homology with a bacterial gene for GAPDH, as we was expected. The amino acid sequence of E. tarda GAPDH (etGAPDH) revealed a <70% similarity with GAPDH proteins in other Enterobacteriaceae. With the application of artificial protein overexpression system in Escherichia coli, the recombinant etGAPDH (rGAPDH) was produced and purified. In this study, Using the purified rGAPDH, the etGAPDH specific polyclonal antibody has been was generated using the purified rGAPDHin this study. The immunoblotting analyses demonstrated that the location of the GAPDH protein is located with the association of is associated with the envelops of E. tarda. The rGAPDH was administrated into Japanese flounder via IP route for evaluation of the protective ability. Although the specific antibody titer against etGAPDH was increased about 3-fold after 4 weeks post-vaccination, the survival rates of vaccinated Japanese flounder and the control group with wild type E. tarda was were 12.5% and 0%, respectively. Our results indicated that rGAPDH is immunoreactive antigen but that it will not generate protective immunity in Japanese flounder.
Keywords
Edwardsiella tarda; vaccine; pathogenesis;
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