Evaluation of Sexing in Boar Sperm Using Chromogenic In Situ Hybridization

Chromogenic In Situ Hybridization을 이용한 돼지 정자의 성 선별

  • Kim, H.H. (Equal contributors) ;
  • Roh, D.E. (Equal contributors) ;
  • Jo, T.K. (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Byun, J.W. (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Lee, J.W. (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Kim, Y.S. (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Hwang, Y.J. (Division of Biological Science, Gachon University of Medicine and Science) ;
  • Kim, D.Y. (Division of Biological Science, Gachon University of Medicine and Science)
  • Published : 2007.09.29

Abstract

돼지 정자의 성 선별에는 일반적으로 유속 세포 분석기를 이용한다. 유속 세포 분석은 DNA량의 차이에 기초하여 정자를 분리하는 기술로써 X 정자와 Y 정자를 90% 정확도로 분리할 수 있다. 그러나 이러한 유속 세포분석 기술은 정자의 손상을 야기해 정자의 기능과 수정능에 영향을 미치므로, 본 연구에서는 특정한 핵산 서열을 탐지할 수 있는 Chromogenic in situ hybridization(CISH)을 그와 비교하여 평가하였다. 유속 세포 분석을 수행하기 위해 정자를 SYBR 14와 PI로 염색하였고, histogram, dotplot, density, contour를 측정하였다. Y 염색체 특이적인 primer를 이용한 PCR로 유속 세포 분석의 정확도를 검사하였다. HRP/DAB 시스템에 기초한 CISH 분석에는 X 또는 Y 염색체에 상보적으로 결합하는 probe가 사용되었다. CISH 분석은 기존의 방법들보다 빠르고 쉬우며 비용이 적게 든다는 장점이 있다. 또한, CISH는 보다 정화한 정자의 선별을 가능하게 하는 것으로 나타났다. 본 연구에 따르면 CISH가 기존의 선별 방법들을 평가하는 기술로서만이 아니라 특정한 성별을 가진 포유동물의 생산에도 사용될 수 있을 것이다.

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