水稻白叶枯病菌hrf2基因表达的His-HarpinXooc蛋白的纯化及分析

摘要

利用已构建的工程菌表达 His-HarpinXooc蛋白,分析 His-HarpinXooc蛋白的纯化条件、产量、等电点和生物活性等。结果表明：1)在使用 HisTrap HP Kit进行纯化时,咪唑浓度对于蛋白纯度影响最大,其中150 mM咪唑最适合于洗脱 His-HarpinXooc蛋白,缓冲液、洗脱次数等对纯化的影响不大；2)每升菌液可获得高纯度的His-HarpinXooc蛋白为20 mg,其等电点约为4.7；3)纯化所得His-HarpinXooc蛋白与不含His标签的HarpinXooc生物活性一致,具有 Harpin蛋白诱导烟草产生过敏反应的典型特征。此为 His-HarpinXooc蛋白的应用奠定基础。%For analyzing the purity,yield,pI and bioactivity,the His-HarpinXooc recombinant protein is expressed from bacteria and purified by HisTrap HP Kit.Through optimizing the conditions,20 mg pure His-HarpinXooc protein can be produced from bacteria suspension per li-ter.The His-HarpinXooc protein is identified by digesting,and the pI of this protein is about 4.7. The Harpin can induce hypersensitive response on tobacco,which is accorded with HarpinXooc . This study is beneficial to the application of His-HarpinXooc protein.