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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
引用本文:刘军,石耀华,尹隽,桂建芳.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[J].遗传学报,2005,32(3):253-263.
作者姓名:刘军  石耀华  尹隽  桂建芳
作者单位:中国科学院水生生物研究所淡水生态与生物技术国家重点实验室武汉发育生物学研究中心,武汉,430072
基金项目:国家自然科学基金重点项目(编号:30130240),国家高技术研究发展计划(863计划)(编号:2001AA2220771),中国科学院创新方向性项目(编号:KXCX2 SW 303)~~
摘    要: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。

关 键 词:银鲫  抑制性差减杂交  差异表达基因  SMART  cDNA  原肠胚  尾芽胚
文章编号:0379-4172(2005)03-0253-11

Screen of Differentially Expressed Genes Between Gastrula Embryos and Tail Bud Embryos in Gynogenetic Gibel Carp (Carassius auratus gibelio)
LIU Jun,SHI Yao-Hua,YIN Jun,GUI Jian-fang.Screen of Differentially Expressed Genes Between Gastrula Embryos and Tail Bud Embryos in Gynogenetic Gibel Carp (Carassius auratus gibelio)[J].Journal of Genetics and Genomics,2005,32(3):253-263.
Authors:LIU Jun  SHI Yao-Hua  YIN Jun  GUI Jian-fang
Abstract: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.
Keywords:gibel carp  suppression subtractive hybridization  differentially expressed gene  SMART cDNA  gastrula embryo  tail bud embryo
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