• 1. Department of Ophthalmology, The 210 PLA Hospital, Dalian 116021, P.R.China;2. Department of Biochemistry and Molecular Biology Of Basic Medicine College,Jilin University, Changchun,Jilin 130021, P.R.China;
Export PDF Favorites Scan Get Citation

【摘要】 目的 觀察不同種培養基中重組人色素上皮衍生因子(rPEDF)融合蛋白的表達。 方法 將前期研究已構建的pET28aPEDF原核表達重組體轉化E.coli BL21大腸桿菌表達宿主菌,酶切鑒定陽性菌落后,分別在M9和LB培養基中用異丙基βD硫代半乳糖(IPTG,IsopropylbetaDthiogalactoside)誘導表達,SDSPAGE電泳檢測表達的PEDF蛋白, 美國ImagePro Plus 分析系統進行蛋白定量分析。結果 LB和M9培養基中均獲得相對分子質量約54×103的rPEDF融合蛋白。但LB培養基獲得的是rPEDF融合蛋白的包涵體,目的蛋白占總蛋白含量為21046%,M9培養基獲得的是可溶性的rPEDF的融合蛋白,目的蛋白占總蛋白含量的1231%。結論 不同種培養基中均有rPEDF 融合蛋白的表達。
【Abstract】 Objective To observe the express of recombinant pigment epithelial derivative facto (rPEDF) in the different medium. Methods The pET28aPEDF was transformed into E.coli BL21. After the colonies were positive identification which were induced by IsopropylbetaDthiogalactoside in medium M9 and LB. The PEDF protein were detected by SDSPAGE and analyzed by American ImagePro Plus system. Results LB and M9 medium obtained the relative molecular mass about 54×103 rPEDF fusion protein. But LB medium obtained the inclusion bodys of rPEDF fusion protein,the purpose protein account for 21.046%;LB medium obtained the soluble rPEDF fusion protein,the purpose protein account for 12.31%. Conclusion The rPEDF protein was expressed in the different medium.

Citation: WANG Yanwei,YANG Nan,FU Weiping,ZHANG Guirong,YANG Hanyi.. The Expression of Recombinant Pigment Epithelium Derived Factor in the Different Medium. West China Medical Journal, 2010, 25(4): 706-. doi: Copy

Copyright ? the editorial department of West China Medical Journal of West China Medical Publisher. All rights reserved