摘要
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(22.7 kD)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。研究结果可为进一步研究PaMsrB1在番木瓜叶绿体中抗氧化机制奠定了基础。
Plant methionine sulfoxide reductase B1（MsrB1） play a critical role in repairing Oxidized chloroplast protein in biotic and abiotic stress. In this study, recombinant papaya MsrB1（PaMsrB1） was successfully expressed in pichia eukaryotic expression system, and purified by Ni2＋-NTA agarose affinity chromatography. western blot revealed that purified recombinant protein was a single protein of the predicted size（22.7 kD）. A protection detection against H2O2-mediated stress and thin layer chromatography assay indicated that recombinant PaMsrB1 has antioxidant capacity and reductase activity in vivo and in vitro, which lay a foundation of further study on the mechanism of PaMsrB1 antioxidant in papaya chloroplasts.
出处
《分子植物育种》
CAS
CSCD
北大核心
2015年第5期1067-1071,共5页
Molecular Plant Breeding
基金
国家自然科学基金(31371918
31171822)资助
关键词
PaMsrB1重组蛋白
表达纯化
抗H2O2氧化
薄层层析
PaMsrB1 recombinant protein
Expression and purification
Against H2O2-mediated stress
Thin layer chromatography
