Proceedings of the Korea Society of Poultry Science Conference (한국가금학회:학술대회논문집)

The Korean Society of Poultry Science (한국가금학회)
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Proteome Analysis of Chicken Embryonic Gonads: Identification of Major Proteins from Cultured Gonadal Primordial Germ Cells

  • Lee, Sang-In (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Han, Beom-Ku (Avicore Biotechnology Institute Inc.) ;
  • Park, Sang-Hyun (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Kim, Tae-Min (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Sin, Sang-Soo (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Lee, Young-Mok (Bioresources Institute, Easy Bio System) ;
  • Kim, Hee-Bal (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Lim, Jeong-Mook (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Han, Jae-Yong (Department of Food and Animal Biotechnology, Seoul National University)
  • Published : 2005.11.18
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Abstract

The domestic chicken (Gallus gallus) is an important model for research in developmental biology because its embryonic development occurs in ovo. To examine the mechanism of embryonic germ cell development, we constructed proteome map of gonadal primordial germ cells (gPGC) from chicken embryonic gonads. Embryonic gonads were collected from 500 embryos at 6 day of incubation, and the gPGC were cultured in vitro until colony formed. After 7-10 days in cultured gPGC colonies were separated from gonadal stroma cells (GSCs). Soluble extracts of cultured gPGCs were then fractionated by two-dimensional gel electrophoresis (pH 4-7). A number of protein spots, including those that displayed significant expression levels, were then identified by use of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and LC-MS/MS. Of the 89 gPGC spots examined, 50 yielded mass spectra that matched avian proteins found in on-line databases. Proteome map of thistype will serve as an important reference for germ cell biology and transgenic research.

배자 생식세포 발달에 관련된 메카니즘을 밝혀내기 위해서, 닭 배자 생식기에서 추출한 원시 생식세포의 단백질체 지도를 만들었다. 총 500 배자를 6일간 배양하여 배자 생식기를 획득했고, 7-10일 배양 후, 배양된 원시생식세포는 2차원 젤 전기 영동법에 의해 분할되어 졌다. 유의적 발현 수준을 나타낸 많은 단백질 스팟 들은 MALDI-TOP 와 LC-MS/MS에 의해 확인되었으며, 89개의 단백질 스팟 중에 50개의 mass spectra 들이 데이터베이스에서 조류 단백질과 일치함을 확인하였다. 본 실험에서 행한 단백질체 지도는 형질전환 연구와 생식세포 생물학 분야에 중요한 참고 문헌으로 가치를 가질수 있을 것이다.

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