水稻抗白叶枯病基因xa5的原核表达及蛋白纯化  

Prokaryotic Expression and Purification of Bacterial Blight Resistance Gene xa5 in Rice

作　　者：范玉龙[1] 张梦诗[1] 李明容[1] 翟文学[2] 夏志辉[1,2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《热带农业科学》2015年第3期36-40,共5页Chinese Journal of Tropical Agriculture

基　　金：国家自然科学基金(No.31071173);国家转基因重大专项(No.2011ZX08001-002);海南省教育厅高校科研资助项目(No.Hjkj2012-12);海南大学科研启动经费(No.KYQD1001)

摘　　要：水稻是重要的粮食作物,其产量高低与病虫害等因素密切相关,由黄单胞杆菌引起的水稻白叶枯病是导致水稻产量减少的主要病害之一。控制白叶枯病最安全、经济有效的方法是培育抗病品种,隐性抗病基因xa5是重要的水稻白叶枯病抗性基因。通过构建显性及隐性xa5基因的原核表达载体(PGEX-4T-1-Xa5和PGEX-4T-1-xa5),转化大肠杆菌BL21,经0.2 mmol/L IPTG 28℃诱导4 h,表达显性Xa5和隐性xa5蛋白;结合western-blotting分析,结果表明表达的蛋白是带有GST标签的融合蛋白,最后利用谷胱甘肽亲和层析柱将蛋白纯化,为后期研究xa5基因的功能提供基础。Rice is an economically important crop, the output of rice is closely related to the factors such as plant diseases and insect pests. Bacterial blight （ BB） caused by Xanthomonas oryzae pv. Oryzae （Xoo） is one of three most destructive diseases in rice which cased output reduce. The cultivation of varieties with high resistance is considered as the most effective and economical option to control this disease. xa5 recessive resistance gene is an important rice bacterial leaf blight resistance genes. In this study, we construct prokaryotic expression vector （PGEX-4T-1-Xa5 and PGEX-4T-1-xa5）, and translate it into BL21, then 0.2 mmol/L IPTG, 28℃, 4 h induced dominant Xa5 and recessive xa5 protein expression. Then in combination with western-blotting analysis, show that the expression of protein is a GST fusion protein and purification of the protein, These results laid a foundation for elucidating characteristic and function of xa5.

关 键 词：水稻 白叶枯病 xa5 原核表达 蛋白纯化 western-blotting 

分 类 号：S511[农业科学—作物学]