花生NBS-LRR类抗病基因的克隆及原核表达  

Cloning and Prokaryotic Expression of Peanut NBS-LRR Resistant Gene

作　　者：刘宇[1,2] 闫彩霞[1] 张廷婷[1] 李春娟[1] 郑奕雄[3] 单世华[1] 

机构地区：[1]山东省花生研究所,山东青岛266100 [2]中国海洋大学,山东青岛266100 [3]仲恺农业工程学院,广州510225

出　　处：《中国农业科技导报》2010年第3期73-78,共6页Journal of Agricultural Science and Technology

基　　金：国家自然基金项目(30771361);国家863计划项目(2006AA10A115);"十一五"国家科技支撑计划项目(2006BAD21B04)资助

摘　　要：根据已知抗病基因的NBS保守区域设计引物,利用PCR技术从花生基因组DNA中克隆得到一个NBS-LRR类抗病基因,序列长539bp,命名为PnAG3。蛋白质序列同源性分析表明多个植物抗病相关蛋白与之有一定的同源性,其中同源性最高的是花生C8_V_253抗性蛋白序列(97%)。将PnAG3基因编码区构建原核表达载体,表达产物分布于包涵体中,大小为47.26KDa,1mmol/LIPTG诱导4h可获得最大表达量。为花生抗黄曲霉品种选育奠定了基础。Based on NBS conserved sequences of the known resistance gene,a pair of primers was designed to be used to amplify a NBS-LRR resistance gene from peanut genome using PCR. The gene was deduced 539 bp long,and named PnAG3. Protein sequence homology analysis showed that the encoded protein PnAG3 had a certain homology with some resistance proteins,among which Arachis cardenasii clone C8_V_253 resistance protein had the highest homology （97%）. A prokaryotic expression vector was constructed to express PnAG3 gene. The product was 47.26 KDa,which was found to be distributed in the inclusion bodies. The maximum prokaryotic expression products would be obtained when induced by 1mmol/L IPTG after 4 h. This study provided a foundation for cultivating anti-Aspergillus flavus peanut varieties.

关 键 词：花生抗病基因 NBS-LRR 生物信息学 原核表达 

分 类 号：S565.2[农业科学—作物学]