摘要
目的:建立DJ-1蛋白原核表达及纯化的技术体系,通过SPR技术评价中药化合物。方法:采用大肠杆菌BL21(DE3)感受态细菌,通过热休克法和IPTG诱导,对GST标记的WT DJ-1蛋白进行原核表达,对GST标签进行切割,采用亲和层析法进行纯化,并通过SPR技术,对7个中药多酚类单体化合物进行WT DJ-1蛋白结合筛选。结果:原核表达并纯化得到高纯度的WT DJ-1蛋白,通过SPR技术筛选出1个与WT DJ-1蛋白有结合的化合物3-O-去甲紫药双口山酮苷(3-O-demethylswertipunicoside,3-ODS)。结论:成功建立了DJ-1蛋白原核表达及纯化的技术体系,可用于评价与DJ-1蛋白结合的化合物。
Objective ： To establish a technical system of the prokaryotic expression and purification of D J- 1 protein for screening polyphenol compounds of traditional Chinese medicine by SPR. Methods： The prokaryotie expression of wild-type （WT） D J-1 was done by heat shock method, and IPTG induction after the plasmid was transformed into the competent bacteria E. coli BL21 （DE3）. Then the GST tag WT DJ-1 protein was purified by glutathione Sepharase 4B affinity chromatography. The purified WT DJ-1 was used to screen 7 polyphenol com- pounds from traditional Chinese medicine by SPR. Results： WT D J-1 protein of high purity was obtained, and one compound, named 3-O-demethylswertipunicoside （3-ODS）, that combined firmly with WT D J-1 protein was screened out. Conclusion： The technical system for the prokaryotic expression and purification of DJ-1 protein has been successfully established. It can be used to screen the compounds that combined with D J-1 protein.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2013年第21期2482-2486,2491,共6页
Chinese Journal of New Drugs
基金
国家"重大新药创制"科技重大专项(2012ZX09103201-042)
方正基金(Founder 201004)
关键词
DJ-1
原核表达
多酚类化合物
中药
SPR
DJ-1
prokaryotic expression
polyphenol compound
traditional Chinese medicine
SPR
