拟南芥VSP1硒代蛋白衍生物的制备和晶体生长

摘要

拟南芥VSP1蛋白是一种具有酸性磷酸酶活性的植物防御蛋白.为利用硒原子的反常散射获取VSP1蛋白晶体X射线衍射的相位信息,以质粒pET-22b为表达载体,大肠杆菌B834(DE3)为宿主菌,在含有硒代甲硫氨酸的M9培养基中诱导表达VSP1硒代蛋白衍生物.通过Ni-NTA亲和层析纯化的目的蛋白经SDS-PAGE检验,纯度在95%以上.通过优化VSP1母体蛋白晶体的生长条件,获得了可衍射的硒代蛋白晶体.%Arabidopsis thaliana VSP1 is a defense protein with acid phosphatase activity. For the use of anomalous X-ray scattering of selenium to provide phase information, VSP1 selenium protein derivatives was expressed using pET-22b in E. Coli B834 in M9 medium containing Se-Met. The target protein was purified using Ni-NTA resin and analyzed by SDS-PAGE. The purity was over 95%. The Se-Met-labeled VSP1 crystals were obtained by optimizing the condition of the native crystals.