小麦抗叶锈基因Lr24候选基因克隆
摘要 【目的】利用同源序列法分离小麦抗病基因同源cDNA序列。【方法】利用抗病基因的保守结构设计引物,从抗叶锈病近等基因系材料TcLr24中扩增出一条703bp的条带RGA1,通过与GenBank比对,选取与RGA1高度同源的若干条带,在它们共有的保守序列位置设计引物,利用cDNA末端快速扩增(RACE)技术扩增抗病同源基因cDNA全长。【结果】扩增到三条全长cDNA,经BLASTp比较,这些序列都含有NBS保守结构域和多个LRR结构域, 与很多已知植物抗病基因的功能相应区域一致。对FRGA-1、FRGA-2和FRGA-3进行荧光定量PCR分析,表明这三个基因在小麦叶片中都是组成型表达。【结论】本研究在小麦材料TcLr24中得到三条抗病基因同源cDNA全长,为研究小麦抗病基因奠定了基础。
Abstract 【Objective】 In this study, resistance gene cDNA homology sequence from wheat was isolated by using homology-based method. 【Method】The primers were designed according to the conserved sequence of resistance gene analogs and a 703 bp fragment was isolated from TcLr24. A few homologous sequences were captured when processing similarity search in GeneBank database. We designed primers based on highly conserved region among the above sequences and the rapid amplification cDNA ends (RACE) was used to obtain the full length sequence of the disease resistance homology gene in the near isogenetic lines TcLr24. 【Result】Three full length cDNA sequences were obtained . BLASTp analysis showed that the deduced amino acids of protein consisted of a NBS conserved domain and many leucine-rich repeats (LRR) domains, which were identical to the conserved domains of many plant resistance genes. These sequences appeared not to be induced by Puccinia triticina and were constitutive genes in the wheat leaf tissue by real time PCR. 【Conclusion】In this study, we obtained three resistance homology sequences which provide the short cut for researching of wheat resistance gene.
张立荣,杨文香,刘大群.小麦NBS-LRR类抗病基因同源cDNA序列的分离与鉴定[J].植物遗传资源学报,2011,12(3):431-436.
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