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Construction, expression, purification and biochemical properties of recombinant Harpin(Xooc)

机译:重组Harpin(Xooc)的构建,表达,纯化及生化特性

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摘要

The Harpins, encoded by the hypersensitive response and pathogenicity (hrp) genes in Gram-negative plant-pathogenic bacteria, can induce a hypersensitive response in non-host plants to provide protection from diseases and pests. To study the structure and function of the Harpins, highly purified Harpin(Xooc) protein is necessary for crystallization. Thus, purification methods for Harpin(Xooc) were established in this study. Harpin(Xooc) was expressed in BL21 cells transformed with a recombinant plasmid, pET-30a-hrf2. Through a series of purification steps, His-select gel column purification and gel-filtration chromatography, highly purified and stable Harpin(Xooc) protein was produced for crystallization. Additionally, the results of the stability of Harpin(Xooc) provided a basis for determination of the optimal crystallization conditions.
机译:Harpins由革兰氏阴性植物致病细菌中的超敏反应和致病性(hrp)基因编码,可以在非寄主植物中诱导超敏反应,从而提供对疾病和害虫的保护。要研究Harpins的结构和功能,高纯度的Harpin(Xooc)蛋白是结晶所必需的。因此,本研究建立了Harpin(Xooc)的纯化方法。 Harpin(Xooc)在重组质粒pET-30a-hrf2转化的BL21细胞中表达。通过一系列纯化步骤,His-select凝胶柱纯化和凝胶过滤色谱法,可以生产出高度纯化且稳定的Harpin(Xooc)蛋白,用于结晶。另外,Harpin(Xooc)的稳定性结果为确定最佳结晶条件提供了基础。

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