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作 者 ：李景景,刘永峰*,郭玉蓉,陈 磊

期 刊 ：西北植物学报 2013年 4期 页码：684~691

关键词：苹果；EIN2；克隆；序列分析；

Keywords:Malus domestica, EIN2, gene cloning, sequence analysis,

摘 要 ：分别以苹果果实总DNA和cDNA为模板,采用PCR、RT-PCR方法扩增、克隆乙烯不敏感基因（ethylene insensitive 2,EIN2）,并利用生物信息学方法分析其核苷酸序列和蛋白质结构。结果表明:（1）以DNA和cDNA为模板的扩增结果完全相同,扩增的EIN2基因片段为4 378 bp,尚未发现有内含子,开放阅读框全长3 282 bp,编码1 093个氨基酸;苹果EIN2相对分子质量为118.9 kD,等电点为5.52,其蛋白可能为脂溶性疏水蛋白。（2）所克隆苹果EIN2基因编码的氨基酸序列与拟南芥(AAD41077.1)、碧桃(ACY78397.1)和葡萄(CAN66374.1)EIN2基因编码的氨基酸序列一致性分别为52%、79%、62%。（3）构建的EIN2基因进化树显示,拟南芥、小盐芥、甜瓜、杨毛果EIN2基因亲缘关系较近,聚为一类;葡萄为一类;蒺藜苜蓿为一类;碧桃、矮牵牛、西红柿聚为一类;苹果单独为一类。而且苹果EIN2基因与碧桃等同源基因的亲缘关系相对较近,与拟南芥、小盐芥同源基因的亲缘关系相对较远。

Abstract:The DNA sequence and cDNA sequence of Malus domestica EIN2 were amplified by PCR and RT-PCR,respectively.Bioinformatic analysis of the nucleotide and sequence of EIN2 was performed.The results showed that:(1)The sequences of M.domestica EIN2 amplified by PCR and RT-PCR both are the same,without intron in it.The M.domestica EIN2 was 4 378 bp,containing one opening reading frame of 3 282 bp and coding 1 093 amino acid residues.The molecular weight of M.domestica EIN2 is 118.9 kD,and its theoretical pI is 5.52,and the protein probably is a liposoluble and lyophobic protein.(2)The multiple sequence alignment indicated that the deduced amino acid sequence of M.domestica EIN2 shared 52%,79% and 62% identity with EIN2 of Arabidopsis thaliana (AAD41077.1),Prunus persica (ACY78397.1) and Vitis vinifera (CAN66374.1),respectively.(3)The constructed phylogenetic tree shows that A.thaliana,Thellungiella halophila,Cucumis melo and Populus trichocarpa were clustered together,while P.persica,Petunia hybrida and Solanum lycopersicum were clustered to another group.V.vinifera,M.domestica and Medicago truncatula were clustered in separate groups,respectively.And M.domestica EIN2 is closely related with the homologous genes of P.persica,et al,and the relationships of it with the homologous EIN2 genes in A.thaliana and T.halophila were remote.

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