摘要
SNX9是近年发现的一种蛋白分选与转运蛋白,属于SNX家族。将原核表达载体pET-SNX9重组质粒转化大肠杆菌BL21(DE3),IPTG诱导表达可获得分子量为70kDa的融合蛋白,Western blotting结果证明此蛋白即为目的蛋白,经检测融合蛋白主要以可溶形式表达。表达产物通过亲和层析和分子筛层析两步纯化,可以获得纯度超过95%的SNX9融合蛋白。分子筛层析中蛋白的出峰体积显示SNX9融合蛋白是以二聚体形式存在。Native-PAGE和动态光散射实验均显示其具有良好的均一性。热稳定性实验表明SNX9蛋白在15℃以下基本稳定。这些信息对进一步研究SNX9的结构和功能具有重要的指导意义。
SNX9 is a novel SNX family protein, which plays important roles in protein sorting and transporting. The prokaryotic expression vector pET-SNX9 construct was transformed into E. coli BL21 （DE3） cell and induced by IPTG. The induced fusion protein （SNX9） was expressed successfully in soluble form. SDS-PAGE indicated that the molecular weight of SNX9 was about 70kDa, and the recombinant protein was confirmed by Western blotting. After affinity and gel filtration chromatography purification, the purity of SNX9 could reach over 95 %. Gel filtration elution volume suggested that SNX9 existed as a dimer in solution. Homogeneity was further confirmed by Native-PAGE and Dynamic Light Scattering （DIS） experiments. In the thermo stability experiment, SNX9 was stable below 15℃ relatively. All these information paved the way for future structural and functional study of SNX9.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2008年第8期11-17,共7页
China Biotechnology
基金
国家“863”计划资助项目(2006AA02Z173)
