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鲤鱼(Cyprinuscarpio)外异蛋白A受体Edar基因的克隆及表达定位

, PP. 99-107

Keywords: 外异蛋白A受体,RACE,整体原位杂交

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Abstract:

外异蛋白A受体(Ectodysplasin-Areceptor,Edar)基因最早是在哺乳动物中发现的,其在皮肤附属物的发育中具有重要的生物学功能。利用Genfishing差异筛选技术对建鲤和镜鲤的皮肤转录表达产物进行筛选,得到Edar基因的部分片段。通过克隆镜鲤mRNA全长发现,镜鲤该基因全长CDS为1389bp,编码一个含有462个氨基酸的蛋白质,包含一个信号肽位点、肿瘤坏死因子受体结合位点序列、跨膜区和死亡结构域。序列比对结果表明,镜鲤Edar蛋白与斑马鱼相似度最高,达88.31%,而与其亲缘关系较远的爪蟾,鸡,人类,小鼠相似度较低,分别是64.88%,63.79%,64.36%,63.50%。对5'-UTR和3'-UTR区进行分析表明,镜鲤与斑马鱼、青鳉5'-UTR的相似度只有23.78%,24.73%,而与非洲爪蟾、鸡、人和老鼠的相似度更低,仅为11.73%、18%、6.50%、10.04%;3'-UTR与青鳉的相似度为35.49%,与非洲爪蟾、人和小鼠的相似度分别为9.42%、8.05%、9.37%。整体原位杂交结果表明,该基因在建鲤和镜鲤鳞片发生时在鳞片着生的皮肤基质中特异表达,而在非鳞片发生区域表达较弱或者不表达,到所有鳞片发育形成后,该基因的表达消失,表明该基因可能参与鲤鱼鳞片的起始发育而不是后期的鳞片模式维持过程。

References

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