红砂肌动蛋白基因片段的克隆及序列分析

Cloning and sequence analysis of Actingene fragment fromReaumuria soongorica

肌动蛋白(Actin)基因是研究植物功能基因表达模式中最常用,也是最重要的内参基因。本研究以泌盐型强旱生植物红砂(Reaumuria soongorica)地上部总RNA为模板,根据其他植物Actin基因的保守序列设计一对简并引物,采用RT-PCR方法克隆其Actin基因片段。序列分析结果表明,该基因片段长度为598bp,编码198个氨基酸,与GenBank中登陆的其他植物Actin基因核苷酸序列和氨基酸序列的一致性分别在82%和90%以上,说明其为Actin基因片段,命名为RsACT。进化分析显示,RsACT与陆地棉(Gossypium hirsutum)和白杨(Populus trichocarpa)肌动蛋白的亲缘关系最近。

Actingene is the most common and important house-keeping gene for analyzing plant functional gene expression patterns.In the present study,the total RNA was extracted from the shoot of salt-secreting super xerophyte Reaumuria soongorica,degenerate primers were designed based on the conserved nucleotide sequences of the Actin genes from other higher plants.Actin gene fragment was obtained by reverse transcription polymerase chain reaction(RT-PCR).The sequence analysis indicated that the Actin gene fragment fromR.soongoricacontained 598 bp,encoding 198 amino acid residues.Multiple alignment with Actingene sequences from other plants in the GenBank showed that Actingene fragment from Reaumuria soongorica had over 82%nucleotide sequence similarity and over 90%amino acid sequence homology with other plants.These results indicated that the cloned gene was Actingene fragment,which was named as RsACT.The phylogenetic tree reconstructed based on the amino acid sequences suggested there might be most closed relationship between RsACT and the actin fromGossypiumhirsutumand Populus trichocarpa.