摘要:
以中国水仙不同花期抑制差减杂交文库(SSH)获得的cDNA片段为基础,采用RACE及RT-PCR技术,分离克隆到1个肌动蛋白基因NtACT1。此基因包含有1个1 134 bp完整阅读框架(ORF),编码377个氨基酸,分子量为41.66 kD,理论等电点为5.31。聚类分析表明:该蛋白与拟南芥的actin4和actin12聚为一类,与桔梗(AEF58501)的亲缘关系最近,其次是杧果(AEO45960)和陆地棉(AAC31886),与登陆的中国水仙actin基因(登录号:AEM89276)亲缘关系较远。半定量RT-PCR分析结果表明:NtACT1在盛花期中国水仙各个器官均有表达,而且表达量基本一致,因此,推断其为组成型表达的肌动蛋白基因。
关键词:
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中国水仙 /
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肌动蛋白 /
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基因克隆 /
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表达
Abstract:
Based on a fragment cDNA from different florescence suppression subtractive hybridization (SSH) of Narcissus tazetta var. chinensis,an actin gene NtACT1 was separated and cloned by RACE and RT-PCR. The NtACT1 cDNA is 1 448 bp,containing a 1 134 bp ORF,encoding a protein of 377 amino acid with a predicted molecular mass of 41.66kD and theoretical isoelectric point of 5.31. Phylogenetic analysis showed that the NtACT1 was clustered together with actin4 and actin12 from which were reproductive actin isovariants. Genetic relationship of NtACT1 is closest with actin of Platycodon grandiflorus (AEF58501),next with actin of Mangifera indica (AEO45960) and Gossypium hirsutum (AAC31886),and farther with actin from Narcissus tazetta var. chinensis (AEM89276). Semi-quantitative RT-PCR revealed that NtACT1 could express at similar level in root,leaf,scape,petal,corolla,stamen and pistil.Therefore,NtACT1 might be constitutive expression actin gene.
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