肾蕨LEAFY基因和启动子的克隆及序列分析  

Cloning and Sequence Analysis of LEAFY Gene and Promoter in Nephrolepis auriculata

作　　者：阚诗卓 徐琪 赵芮 王琦 伍冉 李映 鄢波 Kan Shizhuo;Xu Qi;Zhao Rui;Wang Qi;Wu Ran;Li Ying;Yan Bo(College of Landscape Architecture and Horticulture Sciences,Southwest Forestry University,Kunming,650224)

机构地区：[1]西南林业大学园林学院

出　　处：《分子植物育种》2019年第14期4623-4630,共8页Molecular Plant Breeding

基　　金：云南省高校园林植物与观赏园艺创新团队平台建设项目(31160177)资助

摘　　要：以孢子植物肾蕨叶片为材料,采用RT-PCR、RACE和Tail-PCR相结合的方法克隆得到肾蕨LEAFY(简写为LFY)基因的完整片段,该基因DNA全长为1912bp,包含3个外显子和2个内含子序列,有一个1170bp的完整开放阅读框,编码389个氨基酸。经过对该基因片段的核苷酸序列分析表明,与荚果蕨的LFY基因在结构上有高度的相似性,同源率高达94%。通过Tail-PCR技术克隆得到1521bp的肾蕨LFY基因的启动子序列。用PlantCARE在线启动子预测工具分析表明:该序列含有启动子的特定结构,如TATAbox,CAAT-box等,还有一些其它的调控元件。本研究以肾蕨为材料,通过对肾蕨LFY基因的克隆,为探讨LFY基因在不开花植物的原始功能奠定基础,进一步阐释了LFY基因的结构和功能。Using the leaves of spore plant Nephrolepis auriculata as the material,the complete fragment of LEAFY gene(LFY)from Nephrolepis auriculata was cloned by RT-PCR,RACE and Tail-PCR methods.The fulllength cDNA was 1 912 bp,containing 3 exons and 2 intron sequences.It had a 1 170 bp complete open reading frame,encoding 389 amino acids.Analysis of the nucleotide sequence of the gene fragment showed that it was highly similar to the LFY gene of Matteuccia struthiopteris in structure,and the homology rate was as high as 94%.The promoter sequence of 1 521 bp LFY gene from Nephrolepis auriculata was cloned by Tail-PCR.Analysis with PlantCARE online promoter prediction tool indicated that the sequence contained specific structure of promoter,such as TATA-box,CAAT-box,and other regulatory elements.This study could provide a basis for the analysis of LFY gene expression patterns in ferns,as well as for the molecular biology,genetics,molecular evolution and phylogenetic evolution of ferns.

关 键 词：肾蕨(Nephrolepis auriculata) LFY基因 启动子 克隆 

分 类 号：R73[医药卫生—肿瘤]