Journal of Aquaculture (한국양식학회지)

The Korean Society of Fisheries and Aquatic Science (한국양식학회)
권호 표지

Intraspecies Androgenesis in Mud Loach (Misgurnus mizolepis): I. Optimization of the Egg Inactivation and Haploid Androgenesis Using Transgene Marker

미꾸라지(Misgurnus mizolepis)의 웅성발생성 처녀생식: I. 형질전환 유전자 표지를 이용한 웅성발생성 반수체 유도의 최적화

  • Nam Yoon-Kwon (Department of Aquaculture & Institute of Marine Living Modified Organisms (iMLMO), Pukyong Notional University) ;
  • Noh Choong-Hwan (Marine Resources and Research Department, Korea Ocean Research and Development Institute) ;
  • Kim Dong-Soo (Department of Aquaculture & Institute of Marine Living Modified Organisms (iMLMO), Pukyong Notional University)
  • 남윤권 (부경대학교 양식학과/해양수산형질전환생물연구소) ;
  • 노충환 (한국해양연구원 생물자원본부) ;
  • 김동수 (부경대학교 양식학과/해양수산형질전환생물연구소)
  • Published : 2006.08.01
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Abstract

Induced androgenesis, a form of artificial parthenogenesis is an important tool for the generation and use of genetically isogenic or clonal fish strain. An optimized protocol for the genetic inactivation of mud loach (Misgurnus mizolepis) oocytes (i.e. production of androgenetic haploid) was developed using UV-irradiation. Various dose levels of UV significantly affected the fertilizing capacity of the eggs, hatchability of embryos and incidence of haploidy. Based on the extensive examinations of treatment conditions on embryo viability and haploid incidence, the optimum dose level of UV irradiation was turned out to be $10,800ergs/mm^2$ with 56.9% of hatching success and 94.6% of haploidy. The overall yield of putative androgen under optimized treatment condition was more than 50% out of total eggs inseminated. The success of androgenetic reproduction of haploid genome was verified by flow cytometry and PCR amplification of transgene that is exclusive to either one of parental sexes. However, a small portion $(8\sim11%)$ of presumed androgenetic haploid larvae was proven to contain residual DNA fragment(s) from maternal parent.

동일 종내 미꾸라지(Misgurnus mizolepis) 웅성발생성 인공처녀생식(intraspecific androgenesis)기술을 개발하기 위해 미꾸라지 난 유전물질의 불활성화와 이에 따른 웅성발생성 반수체 유도 조건을 최적화하였다. 다양한 자외선(UV)농도$(0\sim25,650ergs/mm^2)$를 이용하여 미꾸라지 난의 유전학적 불활성화를 시도하였으며 수정률, 부화율 및 반수체 출현율을 평가한 결과 10,800 ergs의 농도에서 가장 우수한 반수체 유도 효율을 보였으며 최초 처리 난 중 50%이상의 웅성발생성 반수체 수율을 확보할 수 있었다. 웅성발생성 개체는 전형적인 haploid syndrome의 특징을 나타내었고 flow cytometry분석 결과 정확히 미꾸라지 반수체(1.4 pg/cell)의 DNA함량을 보유하는 것으로 나타났다. 인위적으로 삽입된 형질전환 유전자 표지를 이용하여 부계 특이적인 인공처녀생식 여부를 확인하였으나 일부 반수체자어들에서$(8\sim11%)$ 모계 유전물질의 잔류가 검출되었다.

Keywords

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