雌核发育银鲫原肠期胚胎和尾芽期胚胎差异表达基因的呈现

Screen of Differentially Expressed Genes Between Gastrula Embryos and Tail Bud Embryos in Gynogenetic Gibel Carp (Carassius auratus gibelio)

构建了雌核发育银鲫原肠期胚胎和尾芽期胚胎间的抑制性差减杂交cDNA质粒文库。对原肠期739个 和尾芽期816个PCR阳性克隆进行斑点杂交,得到72个原肠期和98个尾芽期斑点杂交阳性克隆。测序和基因 数据库比对结果表明:72个原肠期斑点杂交阳性克隆中,包括19个已知基因的cDNA片段和31个没有同源性的 cDNA片段;98个尾芽期斑点杂交阳性克隆中,包括52个已知基因的cDNA片段和37个没有同源性的cDNA片 段。采用虚拟Northern杂交和RT PCR证实了部分基因在银鲫胚胎发育过程中的差异表达。这些差异表达基因 的呈现为进一步研究银鲫胚胎发育的分子机制奠定了基础。

Suppression subtractive hybridization (SSH) cDNA plasmid libraries were constructed between gastrula embryos and tail bud embryos in gynogenetic gibel carp (Carassius auratus gibelio).739 and 816 PCR positive clones were respectively selected to perform dot blot,and 72 dot blot positive clones and 98 dot blot positive clones were obtained from the SSH plasmid libraries specific for gastrula embryos and tail bud embryos.Sequencing analysis and database searches indicated that there were 19 known genes and 31 unknown cDNA fragments in the sequenced 72 dot blot positive clones specific for gastrula embryos,and 52 known genes and 37 unknown cDNA fragments in the sequenced 98 dot blot positive clones specific for tail bud embryos.Moreover,specific expressions of partial genes were further confirmed by virtual Northern blots and RT-PCR.The screen of these differentially expressed genes will help us to understand the molecular mechanism in gibel carp embryogenesis.