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[目的]克隆小白杏(Prunus armeniaca)PaLOX基因全长并对其进行序列分析。[方法]根据已有的ESTs序列,设计3’和5’端RACE引物,利用RT-PCR和RACE技术克隆小白杏PaLOX基因c DNA全长。[结果]PaLOX c DNA全长序列为2 723 bp,含有一个1 002 bp的ORF开放阅读框,编码334个氨基酸,含有PLAT高度保守的特异结构域,预测蛋白质的相对分子质量为26.558 1 k Da,推测理化等电点为8.77,分子式为C1689H2635N459O490S13;该蛋白主要表现为亲水性,α-螺旋和延伸链则散布于整个蛋白质中,空间结构不稳定,无信号肽和跨膜结构,可能存在于线粒体中。进化分析表明,PaLOX氨基酸序列与梅、碧桃、苹果、梨、草莓等蔷薇科聚为一类,相似性依次为98%、99%、78%、77%、63%。[结论]获得了编码小白杏脂氧合酶的基因PaLOX全长序列,登录号为KM067451。
[Objective] The aim of the study was to clone the full-length PaLOX gene of Prunus armeniaca and analyze its sequence. [Method] According to the existing ESTs sequences, 3 ’and 5’ RACE primers were designed and the full length cDNA of PaLOX gene was cloned by RT-PCR and RACE. [Result] The full length sequence of PaLOX cDNA was 2 723 bp and contained a ORF of 1 002 bp encoding 334 amino acids with a highly conserved specific domain of PLAT. The predicted relative molecular mass of the protein was 26.558 1 kDa , Presumed physical and chemical isoelectric point of 8.77, the molecular formula C1689H2635N459O490S13; The protein is mainly manifested as hydrophilic, α-helix and the extension of the chain are scattered throughout the protein, the spatial structure is unstable, no signal peptide and transmembrane structure may exist In mitochondria. Phylogenetic analysis showed that the amino acid sequence of PaLOX clustered together with Rosaceae such as Plum, Petal, apple, pear and strawberry with the similarities being 98%, 99%, 78%, 77% and 63% respectively. [Conclusion] The full-length sequence of PaLOX gene was obtained, encoding accession number KM067451.