Proteome analysis of chicken embryonic gonads: identification of major proteins from cultured gonadal primordial germ cells.
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 (gPGCs) from chicken embryonic gonads. Embryonic gonads were collected from 500 embryos at 6 days of incubation, and the gPGCs were cultured in vitro until colony formed. After 7-10 days in culture, 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 this type will serve as an important reference for germ cell biology and transgenic research.
Han
Beom Ku
BK
Avicore Biotechnology Institute Inc., Hanlim Human Tower, Gunpo City, Gyeonggi-Do, Korea.
Kim
Jin Nam
JN
Shin
Ji Hye
JH
Kim
Jin-Kyoo
JK
Jo
Do Hyun
DH
Kim
Heebal
H
Han
Jae Yong
JY
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005854:Germ Cells / Q000166:cytology / Q000378:metabolism*; D006066:Gonads / Q000166:cytology / Q000196:embryology*; D008297:Male; D020543:Proteome / Q000032:analysis / Q000378:metabolism*
D020543:Proteome