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Molecular probe for identification of cysts of resting cyst of PSP-producer Alexandrium tamarense (Dinophyceae)

분자생물학적 방법을 이용하여 마비성 패류 독소를 생산하는 알렉산드륨 타마렌스 시스트 탐색

  • Cho, Eun-Seob (Harmful Algal Research Division, Author for reprint requests, National Fisheries Research and Development Institute)
  • Published : 2003.04.01

Abstract

Identification of species within the toxin-producing genus Alexandrium is vital for biotoxin monitoring and mitigation decisions regarding shellfish industry. In particular, the discrimination of resting cysts of only A. tamarense from that of Alexandrium spp. is considerable important to fundamentally monitor and predict this species before vegetative cells occur in the nature. Fluorescent cTAM-F1 DNA probe was responsible to not only binding the activity of the vegetative cells in A. tamarense, but also to the resting cysts, which was treated with methanol after fixation and stained by primuline on the surface The location of fluorescence in cultured vegetative cells and resting cysts was almost at tile bottom of the nucleus. The optimal incubation temperature and time using in situ hybridization were 50-$54^{\circ}C$ and 40-60 min, respectively, to penetrate the DNA probe into cell.

알렉산드륨 적조생물 속에서 마비성 패류독소를 생산하는 종을 신속하게 동정하므로 패류양식의 독성 모니터링과 방제에 중요한 역할을 할 수 있다. 자연상태에서 영양세포가 출현하기 전 알렉산드륨 타마렌스의 휴면포자만을 신속하게 분리 동정한다는 것은 근본적인 마비성 패류독소 모니터링 및 예측에 큰 역할을 할 수 있다. cTAM-Fl DNA probe은 알렉산드륨 타마렌스의 영양세포 뿐만 아니라 primuline으로 염색하여 메타놀로 고정한 휴면포자에도 반응이 되었다. 영양세포와 휴면포자에 반응되는 DNA probe 위치는 핵내의 말단 부위에 보였다. DNA probe가 세포내로 삽입되는데 가장 적합한 온도와 시간은 50-$54^{\circ}C$, 40-60분이 좋았다.

Keywords

References

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