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鲤IGF2b基因5'侧翼区序列克隆及不同鲤该区域GC富集区的甲基化分析
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现代农业产业技术体系建设专项资金项目(No.CARS-46),中央级公益性科研院所基本科研业务费专项资金项目(No.2009JBFB01),江苏省自然基金项目(No.BK2010146);


Cloning of IGF2b 5′Flanking Region and Methylation Analysis of Its GC-rich Sequence in Common Carp (Cyprinus carpio)
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    摘要:

    鲤(Cyprinus carpio)胰岛素样生长因子2b(insulin-like growth factor 2b,IGF2b)基因为鲤IGF基因家族重要的成员,已有的研究表明5'侧翼区序列对其功能的发挥有重要作用。因此,采用基因组步移法获取5'侧翼区序列,通过亚硫酸盐修饰的方法分析不同品种该区域GC富集区的甲基化状况。本文成功从鲤基因组DNA中获得695 bp的IGF2b 5'侧翼区序列。已获取的IGF2b 5'侧翼区序列经过TFSEARCH分析后发现了多个转录因子结合位点和TATA框,通过BLAST比对发现,鲤IGF2b 5'侧翼区DNA序列与斑马鱼(Danio rerio)IGF2b基因相比,相似性为87%。检测该区域富含GC的序列17个位点,发现建鲤(C.c.var.jian)有2个个体的3个位点出现甲基化,而黄河鲤(C.c.haematopterus)只有1个个体1个位点出现甲基化,这说明2个品种鲤在这个区域出现甲基化修饰的几率低,也验证了这2品种在该基因该区域GC富集区是稳定的。

    Abstract:

    In Common Carp (Cyprinus carpio) IGF2b is an important member of IGF family, and the 5′ flanking region sequence contributes to its function largely. The 5′ flanking region sequence was cloned by genome walking method and the methylation of GC-rich region in the sequence was also analysed through sulfate-modified method. It was shown that IGF2b 5′ flanking region sequence was 695 bp in length. Multiple transcription factor binding sites and TATA box were found by TFSEARCH analysis. IGF2b 5′ flanking region sequence shared 87% identity with that of zebrafish as revealed by BLAST. Two individuals of Jian Carp(C.c.var.jian)had 3 methylated sites, while only one individual of Huanghe Carp (C.c.haematopterus) had similar modification among the 17 GC-rich regions examined. Thus, methylation modification level is low in this region, and this GC-rich sequence was stable in both Jian Carp and Huanghe Carp.

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张成锋,苏胜彦,朱健,朱文彬,刘珊,董在杰.2012.鲤IGF2b基因5'侧翼区序列克隆及不同鲤该区域GC富集区的甲基化分析.动物学杂志,47(2):98-104.

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  • 收稿日期:2011-09-19
  • 最后修改日期:2012-01-04
  • 在线发布日期: 2012-04-17
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