3个澳洲坚果MYB蛋白靶基因的克隆与生物信息学分析

Cloning and Bioinformatics Analysis of Three MYB Protein Target Genes from Macadamia(Macadamia integrifolia)

【目的】为阐明澳洲坚果MYB(v-myb avian myeloblastosis viral oncogene homolog)蛋白靶基因的功能及分子作用机制提供了科学的理论依据。【方法】基于澳洲坚果转录组数据和RT-PCR方法克隆澳洲坚果MYB蛋白靶基因,并利用生物信息学分析这些基因编码蛋白质的同源性、系统进化、理化性质、亚细胞定位、跨膜域和信号肽。【结果】获得了3个新的澳洲坚果MYB蛋白靶基因MiTOM1、MiTOM2和MiTOM3,GenBank登录号分别为MT319120、MT319121和MT319122。MiTOM1、MiTOM2和MiTOM3都包含典型结构域VHS-ENTH-ANTH,与其它植物来源的TOM蛋白具有极高的序列相似度,与荷花TOM蛋白(XP_010260421.1、XP_010244847.1和XP_010260421.1)的序列相似度分别高达68.93%、75.76%和68.80%。MiTOM1、MiTOM2和MiTOM3蛋白均为亲水性蛋白。MiTOM1可能定位于细胞核,为非分泌蛋白和非跨膜蛋白;MiTOM2可能定位于细胞质,为跨膜分泌蛋白;MiTOM3可能定位于细胞核或微体,为非分泌蛋白和非跨膜蛋白。蛋白二级结构预测显示,MiTOM1、MiTOM2和MiTOM3二级结构主要含有无规则卷曲和α螺旋。【结论】MiTOM1、MiTOM2和MiTOM3包含TOM家族典型结构域VHS-ENTH-ANTH,可能在澳洲坚果体内物质运输及控制形态发育方面起重要作用。

【Objective】This study provides a scientific theoretical basis for elucidating the function and molecular mechanism of macadamia MYB(v-myb avian myeloblastosis viral oncogene homolog)protein target genes.【Method】Based on the macadamia transcriptome data,3 MYB protein target genes of macadamia were cloned by RT-PCR method,and the homology,phylogeny,physicochemical properties,subcellular localization,transmembrane domains,and signal peptides of these genes encoding proteins were analyzed by bioinformatics.【Result】Three new MYB target genes MiTOM1,MiTOM2 and MiTOM3 were cloned.The GenBank accession numbers are MT319120,MT319121 and MT319122 respectively.MiTOM1,MiTOM2 and MiTOM3 which contain the typical domain VHS-ENTH-ANTH,have high sequence similarity with other plant TOM proteins.The amino acid sequence similarity between MiTOM1 and the lotus TOM proteins(XP_010260421.1,XP_01024847.1,XP_010260421.1)reach 68.93%,75.76%,and 68.80%,respectively.MiTOM1,MiTOM2 and MiTOM3 are hydrophilic proteins.MiTOM1 which might be located in the nucleus is non-secretory and non-transmembrane proteins;MiTOM2 which might be located in the cytoplasm is transmembrane secretory proteins;MiTOM3 which might be located in the nucleus or microbodies is non-secretory and non-transmembrane proteins.Protein secondary structure prediction shows that MiTOM1,MiTOM2 and MiTOM3 secondary structures mainly contained irregular crimp andαhelix.【Conclusion】MiTOM1,MiTOM2 and MiTOM3 which include TOM family typical domain VHS-ENTH-ANTH,may play an important role in material transportation and morphological development in macadamia.