番木瓜蛋氨酸亚砜还原酶B1 (PaMsrB1)的真核表达及其活性检测

来源 :第七届生命科学联合学术大会 | 被引量 : 0次 | 上传用户:kof8697
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在生物和非生物逆境下,植物蛋氨酸亚砜还原酶B1 (methionine sulfoxide reductase B1,MsrB1)具有修复叶绿体中被氧化蛋白的功能.本研究首次利用毕赤酵母真核表达系统,胞外分泌表达了番木瓜MsrB1(PaMsrB1)重组蛋白,并通过Ni2+-NTA-琼脂糖亲和层析柱获得了纯化蛋白,western blot结果显示,在预期大小(22.7kDa)出现特异性条带,表明表达纯化的蛋白为PaMsrB1重组蛋白.通过H2O2氧化胁迫和薄层层析实验,从体内和体外证明了获得的重组PaMsrB1具有抗氧化能力和还原酶活性,为进一步研究PaMsrB1在番木瓜叶绿体中抗氧化机制奠定了基础.
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