东乡野生稻RPS2类候选抗病基因的克隆与分析  

Cloning and analysis of candidate RPS2 in Dongxiang wild rice

作　　者：崔玲玲[1,2] 李磊[1,2] 韩小霞[1,2] 邢俊杰[1,2,3] 李玲龙[1,2] 阳志刚[1,2] 罗秀娟[1,2] 李丁[1,2] 谢灵灵[1,2] 曹孟良[1,2,3] 

机构地区：[1]中南大学研究生院隆平分院,湖南长沙410125 [2]湖南西城杂交水稻基因科技有限公司,湖南长沙410205 [3]国家杂交水稻工程技术研究中心,湖南长沙410125

出　　处：《湖南农业大学学报（自然科学版）》2009年第2期111-115,共5页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家“973”项目(2007CB109007);国家“863”重点项目(2006AA100101);国家科技支撑计划项目(2007BAD77B00)

摘　　要：利用NBS保守结构域获得的探针及基于RecA文库富集方法的试剂盒筛选东乡野生稻液体pCAMBIA1301质粒文库,杂交筛选获得阳性克隆,在NCBI上进行BLASTN序列比对后,设计相关引物,扩增获得编码区,并利用生物信息学方法预测其编码蛋白序列,与已克隆的NBS-LRR类抗性基因建立进化树.结果分析表明,其预测蛋白除了部分氨基酸缺失及2个氨基酸不同外,与粳稻大部分序列一致,分子进化树表明其与粳稻、拟南芥RPS2基因的关系最近.Using the probe obtained by use of NBS Conservative Motif on the basis of kit of RecA enrichment, 1 positive clone was screened from the pCAMBIA1301 liquid plasmid library of Dongxiang wild rice by hybridization method.Designed primers were used to amplify the clone based on BLASTN of NCBI. The sequence of protein was predicted and the phylogenetic tree with other NBS-LRR resistance gene was constructed by bioinformatics method. The analysis indicated that sequence of the predicted protein was different from that of japonica rice only in some deletion amino acids and 2 amino acids. The phylogenetic tree indicated that it was closest with RPS2 of japonica rice and Arabidopsis thaliana. The function will be verified by further field identification.

关 键 词：东乡野生稻 液体质粒文库 长片段高保真PCR 生物信息学 

分 类 号：S511.903.4[农业科学—作物学]