XA21和PI-D2激酶蛋白质在酿酒酵母中的表达及其自我磷酸化研究  

Expression and autophosphorylation analysis of XA21 and PI-D2 protein kinases in Saccharomyces cerevisiae

作　　者：王书利[1] 李莉云[1] 尚俊军[2] 王静[2] 刘国振[1] 

机构地区：[1]河北农业大学生命科学学院,保定071001 [2]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《微生物学报》2007年第6期1009-1012,共4页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30328019;30370872;30670175)~~

摘　　要：白叶枯病和稻瘟病是最主要的水稻病害。Xa21是水稻白叶枯病抗性基因,Pi-d2是稻瘟病抗性基因,二者都编码类受体激酶蛋白质。在前期研究中,曾系统地研究了细菌中表达XA21激酶蛋白质的生化活性。在此实验中利用真核表达系统酿酒酵母对Xa21和Pi-d2编码的蛋白激酶进行了表达、纯化及自我磷酸化活性分析,为进一步的生化分析、蛋白质-蛋白质相互作用研究、底物筛选等奠定了基础。Rice bacterial blight and blast are the most crucial rice disease. Xa21 confers resistance to bacterial blight, while Pi-d2 confers resistance to rice blast. Both Xa21 and Pi-d2 encode receptor kinase-like proteins. Biochemical properties of XA21 kinase expressed in bacterial were characterized in our previous report. In this study, both XA21 and PI-D2 kinase domain were PCR amplified and cloned into yeast expression vector pEGH via recombinational cloning strategy, kinase proteins expressed in eukaryotic yeast system was purified and autophosphorylation assay was carried out. The results indicated that XA21 and PI-D2 protein can be detected by SDS-PAGE and showed expected molecular weight. Autophosphorylation assay indicated that yeast expressed XA21 and PI-D2 were active when incubated with P^32 labelled ATP. The experiment provided basic materials for biochemical prosperity analysis, protein-protein interaction and substrate screening research.

关 键 词：水稻 白叶枯病 稻瘟病 抗病基因 酿酒酵母 

分 类 号：S511[农业科学—作物学]