N末端缺失萝卜谷胱甘肽磷脂氢过氧化物酶的表达、纯化及结构预测

Expression,Purification of a Recombinant N-Terminal 32 Amino Acids Deletion Radish PHGPx

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.