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中华大蟾蜍gal-3基因多样性的原因分析

, PP. 171-180

Keywords: 中华大蟾蜍,半乳糖凝聚素3,分子多样性

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Abstract:

使用TAKARA的SMARTerTMRACEcDNAAmplificationKit和改进的QuantscriptRTkit方法制备了3组源于同一中华大蟾蜍(Bufogargarizans)个体皮肤总核糖核酸(RNA)的第一链互补脱氧核糖核酸(cDNA),并以此为模板通过聚合酶链式反应(PCR,polymerasechainreaction)扩增半乳糖凝聚素3(gal-3)的全长开放阅读框(ORF,openreadingframe),然后进行PCR产物的克隆、测序,再通过多种形式的序列比对分析该基因多样性产生的原因.从3组实验得到31个克隆,去除组内和组间重复出现的序列,共获得16个不同的cDNA.相同的序列,相同的单核苷酸多态性位点(singlenucleotidepolymorphism,SNP)以及相同位置相同数目的碱基缺失在3组独立的实验中重复出现,表明gal-3基因多样性.实际上,gal-3基因的多样性导致其编码蛋白的多样性.尽管如此,推导的氨基酸序列比对显示半乳糖凝聚素3糖基结合位点、磷酸化位点以及与其生物学功能关系密切的相关基序均高度保守,暗示中华大蟾蜍皮肤gal-3基因多样性的客观存在.

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