Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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The 16S rDNA Gene Sequencing and Specific Probes Designing for the Identification of Edwardsiella tarda

  • Lee Ju Suk (Division of Pathology, National Fisheries Research and Development Agency) ;
  • Choi Jae Young (Graduate School of Biotechnology, Korea University) ;
  • Sim Doo Saing (Division of Pathology, National Fisheries Research and Development Agency) ;
  • Kim Hyeung Rak (Department of Food science & Biotechnology, Pukyung National University) ;
  • Jung Tae Sung (Department of Microbiology, College of Medicine, Gyeongsang National University) ;
  • Kim Jae Ho (Graduate School of Biotechnology, Korea University) ;
  • Oh Myung Joo (Department of Fish Pathology, Yosu National University)
  • Published : 2000.06.01
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Abstract

DNA probes for the l6S rRNA have been designed for the detection of Edwardsiella tarda. In order to accomplish this purpose, the l6S rRNA gene from E. tarda has been cloned and sequenced. Two highly feasible oligonucleotide probe sites have been determined by the database analysis programs presented by PCGENE and BLAST. These two probes have been evaluated by slot blot hybridization analysis. Hetero- and homo-trimeric templates have been synthesized using these two probe sites. The templates have been further multimerized by PCR to generate between 150 and 300 bp long DIG-11-dUTP labeled probes. Unlike 3' end labeled oligonucleotide probes or templates, multimerized probes showed no cross­hybridization in the given experimental condition. Furthermore, a significant increase in sensitivity has been observed with these probes. This method, we presented here, may be useful for the designing of probes for the detection of other fish pathogenic microorganisms also.

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