小麦RPL21基因同源克隆与表达分析  

CLONING AND EXPRESSION ANALYSIS OF RPL21 GENE IN WHEAT (Triticum aestivum L.)

作　　者：栗现芳[1] 马守才[1] 张改生[1] 牛娜[1] 

机构地区：[1]西北农林科技大学/陕西省作物杂种优势研究与利用重点实验室/小麦育种教育部工程研究中心,陕西杨陵712100

出　　处：《核农学报》2011年第1期6-13,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家863计划重大专项(2009AA101102);国家自然科学基金(31071477);高等学校博士点基金(20090204110024);陕西省13115科技创新工程重大科技专项(2007ZDKG-020);国家杨凌农业生物技术育种中心专项基金(99-1A);西北农林科技大学拔尖人才支持计划项目

摘　　要：为了解60S核糖体蛋白L21(ribosomal protein L21,RPL21)的基因组结构和表达模式,以GenBank数据库中小麦RPL21基因的部分序列为信息探针,采用RT-PCR和PCR技术从小麦多子房株系幼穗中克隆了RPL21基因的cDNA与DNA片段,并对该基因在小麦多子房近等基因系间的表达差异和多子房株系中的时空表达模式进行了分析。结果表明,所克隆的小麦RPL21基因cDNA序列(GenBank登录号:HM138480)长521bp,编码164个氨基酸;DNA序列(GenBank登录号:HM138481)长1600bp,含有2个外显子和1个内含子。半定量RT-PCR分析显示,该基因在多子房株系幼穗中表达少量上调;在多子房株系不同时期幼穗中的表达量随着幼穗的发育呈上升趋势;不同组织中具体表达模式为茎顶端≈幼根>幼穗>幼叶,生长旺盛部位表达量较高。To understand the genome structure and expression patterns of 60S ribosomal protein L21（RPL21）,partial sequence of RPL21 gene from the GenBank database was used as a query probe to blast the GenBank database.Based on the assembled homologous cDNA sequence,both cDNA and DNA sequences of RPL21 from multi-ovary lines were isolated and characterized by RT-PCR.Furthermore,differential expression of genes between the multi-ovary line and mono-ovary line,expression patterns of genes in different development stages of ears and different tissues of multi-ovary line were analyzed by semi-quantitative RT-PCR.The cDNA sequence（GenBank accession No.HM138480） was 521bp in length and the open reading frame encoded a peptide of 164 amino acids.The DNA sequence（GenBank accession No.HM063421） was 1600bp in length,which contained two exons and one intron.According to the expression analysis,the expression of the gene in 2 ～ 4mm ears of multi-ovary line was higher than that in mono-ovary line.This gene was upregulated with the development of young ear of multi-ovary line,and arrived at the highest expression levels at 8 ～ 9mm young ear stage.It was speculated that more RPL21 were needed for the growth and development of floral organ.Besides,the expression patterns of the gene in different tissues of multi-ovary lines were in the order of stem shoot meristem ≈ young root 〉young ear 〉young leaf,indicating that they were abundantly expressed in various tissues,expression levels of RPL21 genes was the highest in rapidly growing tissues.

关 键 词：多子房小麦 RPL21基因 克隆 表达分析 

分 类 号：S512.1[农业科学—作物学]