硫氧还蛋白融合表达系统表达的小鼠内皮抑素之纯化和生物学活性鉴定
蛋白,,内皮抑素;硫氧还蛋白融合表达系统;融合蛋白,0引言,1材料和方法,2结果,【参考文献】
Purification of mouse endostatin in thioredoxin fusion expression system and identification of its biological activityMA WeiLie,LIU Fang,HE ChengWei,HE ZhenHui
Department of Biochemistry,Guangdong Medical College,Zhanjiang 524023,China
【Abstract】 AIM:To purify mouse endostatin by using thioredoxin fusion expression system and to identify its biological activity. METHODS: The pThioHisendo recombinant plasmid was transformed into BL21. After induction with IPTG, thioredoxinendo fusion protein was expressed in BL21 and the product was identified as inclusion body by SDSPAGE. The expression product was purified by affinity chromatography through Nicolumn. The purified protein was detected by chicken chorioallantoic membrane(CAM) angiogenesis inhibitory assay and endothelial cell proliferation inhibitory assay. RESULTS: High purity thioredoxinendostatin fusion protein was obtained by SDSPAGE analysis. The purity of the protein was over 95%. The protein possessed the inhibitory activity of endothelial cell proliferation and inhibited the angiogenesis of CAM [inhibitory rate: (49.6±4.1)%, (53.2±2.3)%, (55.0±3.6)% and (67.1±5.7)%]. CONCLUSION: The mouse endostatin recombinant fusion protein expressed in E.coli by using thioredoxin fusion expression system is easy to be purified and possesses high activity. ......
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