鹌鹑gcl基因BTB/POZ结构域编码序列的克隆及组织中的表达

Germ cell-less(GCL)是与原始生殖细胞发生相关的重要因子,果蝇、斑马鱼、青鳉鱼和小鼠的GCL蛋白都含有一个进化上保守的BTB/POZ结构域。本研究应用简并PCR克隆技术,在鹌鹑(Coturnix coturnix)中扩增得到319bp的gcl基因(包含BTB/POZ结构域编码序列)保守序列。通过与线虫、果蝇、斑马鱼、小鼠和人gcl同源序列的比对,发现该片段与它们的同源性分别为52.4%、55.4%、84.6%、79.6%和76.8%。采用RT-PCR和mRNA整体原位杂交方法,研究了gcl基因在不同组织和原条期胚胎中的表达。结果显示,该基因在卵巢、精巢和肝中表达;gcl mRNA存在于原条期胚胎的生殖新月区。

Cloning and Expression of Quail gcl BTB/POZ Domain Coding Sequence

Germ cell-less (GCL) plays an essential role in the formation of primordial germ cells.An evolutionally conserved domain,BTB/POZ domain,was found in the GCL homologue proteins of Drosophila,zebrafish,medaka and mouse.A gcl conserved sequence of 319 bp,which contained BTB/POZ domain coding sequence,was isolated from Quail (Coturnix coturnix) genome DNA using the BTB degenerated primers.The conserved sequence of Quail gcl showed 52.4%,55.4%, 84.6%, 79.6% and 76.8% identity to Caenorbabditis elegan,Drosophila,zebrafish,mouse and human gcl homologous fragment respectively.RT-PCR revealed its expression in ovary,testis and liver.By whole-mount in situ hybridization,gcl mRNA was detected in the germinal crescent of primitive streak stage in Quail embryo.