芒果乙烯受体ETR1和ERS1基因的克隆及序列分析

Cloning and sequence analysis of ethylene receptor ETR1 and ERS1 genes from mango(Mangnifera indica L.)

  • 摘要: 目的克隆芒果乙烯受体ETR1和ERS1基因(MiETR1和MiERS1),并进行生物信息学分析,为研究乙烯受体在芒果成熟和衰老过程中的调控机制提供理论参考.方法以芒果为材料,应用RACE技术扩增MiETR1和MiERS1基因的cDNA序列,并应用生物信息学方法对其编码的氨基酸序列进行预测分析.结果克隆获得的MiETR1基因cDNA序列全长2570 bp,开放阅读框(ORF)2220 bp,编码739个氨基酸;MiERS1基因的cDNA序列全长2171 bp,ORF 1890 bp,编码629个氨基酸;GenBank登录号分别为KY002681和KY002682.MiETR1和MiERS1蛋白均为含跨膜结构的非分泌型疏水蛋白,属于乙烯受体ETR1家族成员,以Ser为主、Thr为辅的磷酸化修饰调控乙烯信号转导,均含有3个典型的模块,即GAF结构域、HisKA结构域和HATPase_c结构域,MiERS1蛋白较MiETR1缺少REC结构域.不同物种ETR1和ERS1蛋白的氨基酸序列具有高度保守性,MiETR1和MiERS1均与甜橙(Citrus sinensis)和克莱门柚(Cit-rus clementina)的ETR1和ERS1蛋白亲缘关系较近,但二者分别聚在两个不同的分支上,表明两个蛋白存在一定的遗传分化.结论乙烯受体基因在芒果成熟和衰老过程中发挥调控作用.

     

    Abstract: ObjectiveEthylene receptor ETR1 and ERS1 genes(MiETR1 and MiERS1)from mango(Mangnifera in-dica L.)were cloned,and bioinformatics analysis was conducted in order to provide theoretical reference for further study of functional mechanism of the ethylene receptors in mango ripening and senescencing.MethodUsing mango fruit as material,full-length cDNA sequences of MiETR1 and MiERS1 by RACE technology was amplified,and the encoded ami-no acid sequences were predicted by bioinformatics method.ResultThe full-length cDNA sequence of cloned gene Mi-ETR1 was 2570 bp,an open reading frame(ORF)of 2220 bp,encoding 739 amino acids. The full-length cDNA sequence of cloned gene MiERS1 was 2171 bp,and it was consisted of an ORF of 1890 bp,encoding 629 amino acids. The Gen-Bank accession numbers for the two were KY002681 and KY002682. Bioinformatic analysis showed that the MiETR1 and MiERS1 proteins were non secreted hydrophobic protein with transmembrane domain,belonging to the ETR1 family member of ethylene receptor with Ser-based,Thr-assisted phosphorylation-modulatory. MiETR1 and MiERS1 proteins contained three typical modules,namely GAF domain,HisKA domain and HATPase_c domain. MiERS1 protein lacked REC domain compared with MiETR1. The amino acid sequences of ETR1 and ERS1 proteins in different species were highly conserved. MiETR1 and MiERS1 had high homologous with the ETR1 and ERS1 proteins of Citrus sinensis and Citrus clementina. However,the two clustered on two different branches,indicating that there was genetic differentiation between the two proteins at certain extents.ConclusionEthylene receptor gene plays a regulatory role in mango fruit maturation and senescence.

     

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