摘要
将N末端定位信号缺失萝卜PHGPx(RsPHGPx)的cDNA片段克隆到表达载体pGEX-6P-1上,并转化至大肠杆菌内进行表达.通过GST亲和层析、离子交换层析和凝胶过滤层析,制备了用于晶体学研究的RsPHGPx.其浓度约为10 g/L,纯度超过95%,具有PHGPx活性.三维结构同源建模显示RsPHGPx的结构为典型的硫氧还蛋白折叠形式.
An N-terminal 32 amino acids radish PHGPx gene (△32RsPHGPx) was cloned into expression vector pGEX-6P-1 and transformed into E.coli strain BL2.1 (DE3). A32RsPHGPx for crystallization was prepared via Glutathione SepharoseTM affinity, cation-exchange and gel filtration chromatographies. The concentration of it was 10 g/L and the purity was over 95%. △32RsPHGPx showed obvious PHGPx activity towards lipid hydroperoxides. In addition, a tertiary structure model of △32RsPHGPx displayed the thioredoxin fold.
出处
《吉林大学学报(理学版)》
CAS
CSCD
北大核心
2008年第1期148-153,共6页
Journal of Jilin University:Science Edition
基金
国家自然科学基金(批准号:3017008039770078)
国家重点基础研究发展计划项目基金(批准号:2006CB101703)
国家863计划项目基金(批准号:2007AA100604)
清华-裕元医学科学研究基金
关键词
萝卜
谷胱甘肽磷脂氢过氧化物酶
表达
纯化
radish
phospholipid hydroperoxide glutathione peroxidase
expression
purification