抗禾谷孢囊线虫基因中核苷酸结合位点区(NBS)-亮氨酸重复序列区(LRR)的克隆与序列分析  

Cloning and Sequencing of Nucleotide-binding Site (NBS)-Leucine-rich Repeats (LRR) Region of Cereal Cyst Nematode Resistance Gene

作　　者：刘毅[1] 翟旭光[1] 吴芳[1] 邓光兵[1] 潘志芬[1] 余懋群[1] 

机构地区：[1]中国科学院成都生物研究所

出　　处：《应用与环境生物学报》2006年第6期745-749,共5页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金资助项目(No.30471150);国家"863"计划项目(2003AA207100);中国科学院知识创新工程重要方向性项目(KSCX2-SW-304);四川省作物育种攻关项目资助~~

摘　　要：根据源于节节麦抗禾谷孢囊线虫基因Cre3及已经克隆的NBS-LRR类植物抗病基因的NBS与LRR区保守序列分别设计特异引物,从易变山羊草基因组中PCR扩增得到两个扩增条带,它们的大小分别约为530bp和1200bp.经克隆测序发现,这两个序列分别长为532bp和1175bp,且是连续的.它们有32bp的共同序列,总长为1675bp,包含了一个NBS-LRR区和一个不完整的开放阅读框,没有起始密码子、终止密码子和内含子结构.它编码一个557个氨基酸的蛋白质,分子量(Mr)为63.537×103,含有NBS区保守模体ILDD,ESKILVTTRSK,KGSPLAARTVGG,RRC-FAYCS,EGF,以及LRR区的保守模体aXXLXXLXXL.它与Cre3的核苷酸和氨基酸同源性分别为87.8%和77%,可能是一个抗禾谷孢囊线虫基因.According to the conversed sequences of NBS and LRR region of cereal cyst nematode resistance gene Cre3 from wild wheat （ Triticum tauschlii） and the known NBS - LRR group resistance genes, two pairs of specific primers were designed for NBS and LRR region, respectively. Through PCR, one band of approximately 530 bp was amplified by the specific primers for conversed NBS region and another band of approximately 1 200 bp was amplified by the specific primers for conversed LRR region. After sequencing, two sequences had 1 675 bp in total with a 32 bp common nucleotide sequence, which was registered as RCCN in the GenBank. RCCN contained a NBS - LRR domain and an incomplete open reading frame without initiation codon, terminator codon and intron. Its exon encoded a peptide of 557 amino acid residues. The molecular weight of the protein from the amino acid was 63. 537 x 103. The amino acid sequence of RCCN contained conserved motifs： ILDD, ESKILVTTR- SK, KGSPLAARTVGG, RRCFAYCS, EGF,and aXXLXXLXXL, which belonged to the conserved motif of LRR region. RCCN shared 87.8% nucleotide sequence and 77% amino acid sequence identity with cereal cyst nematode gene Cre3. It might be a novel cereal cyst nematode resistance gene. Fig 6, Ref 13

关 键 词：易变山羊草 抗禾谷孢囊线虫基因 核苷酸结合位点区-亮氨酸重复序列区 克隆和序列分析 

分 类 号：S432.45[农业科学—植物病理学]