Molecular Characterization of Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) Gene of Field Isolate of Edwardsiella tarda

Megha K. Bedekar, Praveena K. Soman, K. V. Rajendran, Gayatri Tripathi, M. Makesh

Aquatic Animal Health, Central Institute of Fisheries Education (ICAR), Mumbai, India.

DOI: 10.4236/abb.2015.62009                

Abstract

The E. tarda bacterial culture isolated from infected fish, was confirmed by morphological and biochemical characterization. GAPDH gene of 996 bp was amplified from bacterial genomic DNA. GAPDH gene was cloned in pTZ57R/T cloning vector. The positive clone was sequenced. The sequencing result showed very homology with published sequence from pathogenic E. tarda. The sequence (nucleotide and amino acid) divergence values were very low between E. tarda isolates from India (KF142190) and China (HQ697337-38) (Figure 4 and Figure 5). However, the divergence value was high when compared with E. tarda isolates from Japan (AB198939) and this value was higher than the inter-specific divergence value (E. tarda-E. ictaluri, E. tarda-E. coli & E. tarda-Klebsiella species).

Keywords

E. tarda, GAPDH Gene, Vaccine, Cloning

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Hanneson, R. (2003) Aquaculture and Fisheries. Marine Policy, 27, 169-178. http://dx.doi.org/10.1016/S0308-597X(02)00083-0
[2]	Leong, J.C. and Fryer, J.L. (1993) Viral Vaccines for Aquaculture. Annual Review of Fish Diseases, 3, 225-240. http://dx.doi.org/10.1016/0959-8030(93)90036-B
[3]	Kawai, K., Liu, Y., Ohnishi, K. and Ohnishi, S. (2004) A Conserved 37 kDa Outer Membrane Protein of Edwardsiella tarda Is an Effective Vaccine Candidate. Vaccine, 22, 3411-3418. http://dx.doi.org/10.1016/j.vaccine.2004.02.026
[4]	Bolton, A., Song, X.M., Willson, P., Fontaine, M.C., Potter, A.A. and Perez-Casal, J. (2004) Use of the Surface Proteins GapC and Mig of Streptococcus dysgalactiae as Potential Protective Antigens against Bovine Mastitis. Canadian Journal of Microbiology, 50, 423-432. http://dx.doi.org/10.1139/w04-016
[5]	Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. and Kumar, S. (2011) MEGA5: Molecular Evolutionary Genetics Analysis Using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution, 28, 2731-2739. http://dx.doi.org/10.1093/molbev/msr121
[6]	Park, S.B., Aoki, T. and Jung, T.S. (2012) Pathogenesis of and Strategies for Preventing Edwardsiella tarda Infection in Fish. Veterinary Research, 43, 67. http://dx.doi.org/10.1186/1297-9716-43-67
[7]	Liu, Y., Oshima, S. and Kurohara, K. (2005) Vaccine Efficacy of Recombinant GAPDH of Edwardsiella tarda against Edwardsiellosis. Microbiology and Immunology, 49, 605-612. http://dx.doi.org/10.1111/j.1348-0421.2005.tb03652.x