砂藓丙二烯氧化环化酶基因RcAOC1的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Allene Oxide Cyclase Gene RcAOC1 in Racomitrium canescens

本研究采用PCR技术从砂藓（Racomitrium canescens）中克隆得到丙二烯氧化环化酶（allene oxide cyclase,AOC）基因,命名为RcAOC1。构建了RcAOC1过表达转基因烟草（Nicotiana tabacum）并对T0代转基因烟草进行了基因型鉴定。对RcAOC1进行了相关生物信息学分析,RcAOC1的cDNA开放阅读框长561 bp,编码含有186个氨基酸的多肽序列。RcAOC1蛋白理论等电点为5.16;不稳定系数为56.25,属于不稳定的蛋白质;脂肪指数为25.13,属于疏水性蛋白质。对蛋白的跨膜结构域进行分析,结果显示RcAOC1基因编码的蛋白不含跨膜区。预测RcAOC1蛋白不含有信号肽,属于非分泌型蛋白。砂藓中AOC1蛋白与其它植物中的AOC蛋白质序列具有较高的相似性,其中与小立碗藓AOC蛋白亲缘关系最近。本研究的结果为进一步研究RcAOC1基因的应用奠定了理论基础。

In this research, an allene oxide cyclase（AOC） gene was cloned from Racomitrium canescens, named RcAOC1. RcAOC1 overexpression transgenic tobacco（Nicotiana tabacum） was constructed and genotypic identification of T0 generation transgenic tobacco was carried out. The related bioinformatics analysis of RcAOC1 was carried out. The length of RcAOC1 cDNA open reading frame（ORF） was 561 bp, encoding a polypeptide sequence containing 186 amino acids. For RcAOC1, theoretical isoelectric point was 5.16 and the unstable coefficient was56.25 belonging to the instable protein; The fat index was 25.13 belonging to the hydrophobic protein. Transmembrane domain of RcAOC1 was analyzed and the results showed that RcAOC1 contained no transmembrane area. RcAOC1 protein was predicted with no signal peptides, indicating that it was a non-secretory protein.RcAOC1 protein showed high similarity with AOC proteins of other plant species in sequences, among which Physcomitrella patens AOC protein performed closest relationship with RcAOC1. The results of this study might provide theoretical foundation for further studies on the application of RcAOC1.