欧李ChIRT1基因的克隆与生物信息学分析

Cloning and Bioinformatics Analysis of ChIRT1 Gene in Cerasus humilis

欧李(Cerasus humilis)是蔷薇科矮生灌木,果实中的钙、铁等营养物质丰富。铁调转运基因1(IRT1)编码的蛋白是ZIP金属转运家族中主要的铁转运蛋白,负责植物中金属离子的转运,尤其是铁的吸收。本研究从欧李叶片中提取RNA,逆转录得到cDNA;参照小金海棠(Malus xiaojinensis)MxIRT1基因设计引物;克隆了欧李ChIRT1基因,并进行了生物信息学分析。结果表明,ChIRT1全长1417 bp,含有1个678 bp的开放阅读框,编码226个氨基酸。推测ChIRT1蛋白的分子量约为24 kD,理论等电点为7.72,含有4个跨膜结构域,属于跨膜蛋白。ChIRT1含有一个ZIP结构域,二级结构中α-螺旋占44.69%,无规则卷曲占38.94%。系统发育分析表明,欧李的ChIRT1基因与蔷薇科的IRT1基因关系密切,与樱桃李(Prunus persica)的PpIRT1基因相似度最高。本研究克隆了欧李ChIRT1基因,并分析了其序列特征,为研究欧李ChIRT1基因功能及铁调节机制提供了依据。

Cerasus humilis is a dwarf shrubby fruit tree belonging to the Rosaceae family with a lot of calcium,iron and other nutrients in the fruit contains.Iron-regulated transporter 1(IRT1)is the main iron transporter of the ZIP metal transporter family,responsible for the transport of metal ions in plants,especially the absorption of iron.In this study,RNA was extracted from the leaves of Cerasus humilis and cDNA was obtained by reverse transcription.The primer was designed with reference to the IRT1 gene sequence of Malus xiaojinensis.ChIRT1 gene was cloned and bioinformatics analyzed.The results showed that the total length of ChIRT1 was 1417 bp,with an open reading frame of 678 bp,and encoded 226 amino acids.It is estimated that the molecular weight of ChIRT1 was about 24 kD and a theoretical isoelectric point was 7.72.Ch IRT1 contained four transmembrane domains and belonged to transmembrane proteins.ChIRT1 protein contained a ZIP domain,with alpha helix accounts for44.69%and random coil accounting for 38.94%of secondary structure.Phylogenetic analysis showed that ChIRT1 of Cerasus humilis was closely related to IRT1 of rosaceae plants,and had the highest similarity with PpIRT1 of Prunus persica.The ChIRT1 gene was cloned and its sequence characteristic was preliminarily analyzed in this study,which provided the research basis for the Ch IRT1 function and iron regulation mechanism of Cerasus humilis.