Cloning and Prokaryotic Expression of Peanut NBS-LRR Resistant Gene

doi:10.3969/j.issn.1008-0864.2010.03.13

›› 2010, Vol. 12 ›› Issue (3): 73-78.DOI: 10.3969/j.issn.1008-0864.2010.03.13

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Cloning and Prokaryotic Expression of Peanut NBS-LRR Resistant Gene

LIU Yu1,2, YAN Cai-xia1, ZHANG Ting-ting1, LI Chun-juan1, ZHENG Yi-xiong3, SHAN Shi-hua1

(1.Shandong Peanut Research Institute, Shandong Qingdao 266100|2.Ocean University of China, Shandong Qingdao 266100|3.Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China)

Received:2009-12-17 Revised:2010-04-06 Online:2010-06-15 Published:2010-05-31

花生NBS-LRR类抗病基因的克隆及原核表达

刘宇1,2,闫彩霞1,张廷婷1,李春娟1,郑奕雄3,单世华1

(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)

通讯作者: 单世华,研究员,博士,主要从事花生种质资源创新利用研究。E-mail:shhshan@sina.com
作者简介:刘宇,硕士研究生,主要从事食品安全与控制研究。E-mail:shishui87@163.com。
基金资助:
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。

Abstract

Abstract:

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.

Key words: peanut disease resistant gene, NBS-LRR, bioinformatics, prokaryotic expression

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539 bp,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47.26 KDa,1 mmol/L IPTG诱导4 h可获得最大表达量。为花生抗黄曲霉品种选育奠定了基础。

关键词: 花生抗病基因;NBS-LRR;生物信息学;原核表达

CLC Number:

S565.2

LIU Yu1,2, YAN Cai-xia1, ZHANG Ting-ting1, LI Chun-juan1, . Cloning and Prokaryotic Expression of Peanut NBS-LRR Resistant Gene[J]. , 2010, 12(3): 73-78.

刘宇1,2,闫彩霞1,张廷婷1,李春娟1,郑奕雄3,单世华1. 花生NBS-LRR类抗病基因的克隆及原核表达[J]. , 2010, 12(3): 73-78.

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Cloning and Prokaryotic Expression of Peanut NBS-LRR Resistant Gene

花生NBS-LRR类抗病基因的克隆及原核表达

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