CAJ | 학술논문

为获得番茄抗晚疫病广谱性基因信息,采用同源电子克隆法,基于番茄蛋白序列数据库,以马铃薯晚疫病广谱抗性蛋白 Rpi-blb1为种子序列,获得番茄疾病抗性蛋白 RGA4,并进行基因电子定位和基因结构、模体、二级结构、基因进化及基因电子表达等分析,以证明两者的进化关系.结果表明：番茄 RGA4蛋白(XP_004245923．1)序列具有 NB-ARC 和 LRR8两个保守结构域,定位于第8条染色体 SL2．40区间,相应基因位于8号染色体序列的57228847－57232935 bp 区间,长度为4089 bp,由2个外显子和1个内含子组成,编码988个氨基酸序列,该蛋白为不稳定分泌球形蛋白；番茄 RGA4蛋白和马铃薯 Rpi-blb1蛋白在二级结构分布、基因序列组成、基因定位等方面相似性较高,可确定两者为垂直同源关系,但在基因进化和模体组成方面存在差异,可能导致两者功能上的不同.研究结果可为番茄晚疫病广谱抗性基因克隆及其利用提供理论依据.
위획득번가항만역병엄보성기인신식,채용동원전자극륭법,기우번가단백서렬수거고,이마령서만역병엄보항성단백 Rpi-blb1위충자서렬,획득번가질병항성단백 RGA4,병진행기인전자정위화기인결구、모체、이급결구、기인진화급기인전자표체등분석,이증명량자적진화관계.결과표명：번가 RGA4단백(XP_004245923．1)서렬구유 NB-ARC 화 LRR8량개보수결구역,정위우제8조염색체 SL2．40구간,상응기인위우8호염색체서렬적57228847－57232935 bp 구간,장도위4089 bp,유2개외현자화1개내함자조성,편마988개안기산서렬,해단백위불은정분비구형단백；번가 RGA4단백화마령서 Rpi-blb1단백재이급결구분포、기인서렬조성、기인정위등방면상사성교고,가학정량자위수직동원관계,단재기인진화화모체조성방면존재차이,가능도치량자공능상적불동.연구결과가위번가만역병엄보항성기인극륭급기이용제공이론의거.
Using electronic cloning method,the putative disease resistance protein RGA4 and the homologue of the potato Rpi-blb1protein were obtained from tomato’s protein sequences database in NCBI by protein-blast when the sequence of late blight resistant protein Rpi-blb1 from potato was used as query sequence,then,they were compared in gene mapping,gene structure,modify,secondary structure,topological structure and electronic gene expression pro-file,etc,for proving their evolutionary relationship.The result showed that tomato RGA4 protein (XP_004245923.1) was composed of 988 amino acid residues with NB-ARC and LRR8 conserved domain,the corresponding cDNA se-quence (XM-004245875.1)was located on SL2.40 region in the chromosome 8,with genomic DNA length 4 089 bp, the alignment region in chromosome 8 between 57228847 - 57232935 bp,and composed of two exons and one intron.The tomato RGA4 protein was an unstable globular secretory protein without transmembrane structure.To-mato RGA4 protein was regarded as an orthologous of potato Rpi-blb1 protein because they were similar in secondary structure composition,gene sequence composition,gene mapping,etc,but their minor differences in gene evolution and composition could result in different functions.This study would provide theory for cloning late blight resistant gene and its utilization in tomato breeding of disease resistance.