Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Diversity of Marine Microbes by PCR-DGGE

PCR-DGGE를 이용한 해양미생물의 다양성 조사

  • Kim, Yeong-Jin (Department of Microbiology and Immunology, College of Medicine, Pusan National University) ;
  • Cho, Hyo-Jin (Department of Microbiology and Immunology, College of Medicine, Pusan National University) ;
  • Yu, Sun-Nyoung (Department of Microbiology and Immunology, College of Medicine, Pusan National University) ;
  • Kim, Kwang-Youn (Department of Microbiology and Immunology, College of Medicine, Pusan National University) ;
  • Kim, Hyeung-Rak (Division of Food Science and Biotechnology, Pukyung National University) ;
  • Ahn, Soon-Cheol (Department of Microbiology and Immunology, College of Medicine, Pusan National University)
  • 김영진 (부산대학교 의과대학 미생물학 및 면역학 교실) ;
  • 조효진 (부산대학교 의과대학 미생물학 및 면역학 교실) ;
  • 유선녕 (부산대학교 의과대학 미생물학 및 면역학 교실) ;
  • 김광연 (부산대학교 의과대학 미생물학 및 면역학 교실) ;
  • 김형락 (부경대학교 수산과학대학 식품생명과학부) ;
  • 안순철 (부산대학교 의과대학 미생물학 및 면역학 교실)
  • Published : 2007.12.31
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Abstract

Recently, the development of various culture-independent identification techniques for environmental microbes has greatly enhanced our knowledge of microbial diversity. In particular, denaturing gradient gel electrophoresis (DGGE) of 16S rDNA fragments, generated using the polymerase chain reaction (PCR) is frequently used to examine the diversity of environmental bacterial populations. This method consists of direct extraction of the environmental DNA, amplification of the 200-600 bp 16S rDNA fragments with universal primers, and separation of the fragments according to their melting point on a denaturing gradient gel. In this study, we investigated the seaside microbial community in coastal areas of Busan, Korea, using culture-independent techniques. First, marine genomic DNA was extracted from seawater samples collected at Songjeong, Gwangahn, and Songdo Beaches. Then, PCR was used to amplify the bacterial 16S rDNA using universal primers, and DGGE was used to separate the amplified 500 bp 16S rDNA fragments. Finally, the tested 16S rDNA genes were further analyzed by sequencing. Based on these experiments, we found that DGGE analysis clearly showed variation among the regional groups. It can be used to monitor rapid changes in the bacterial diversity of various environments. In addition, the sequence analysis indicated the existence of many unculturable bacteria, in addition to Arcobacter, Pseudoaltermonas, and Vibrio species.

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References

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