不同培养基中重组人色素上皮衍生因子融合蛋白的表达

目的观察不同种培养基中重组人色素上皮衍生因子（rPEDF）融合蛋白的表达。方法将前期研究已构建的pET28a-PEDF原核表达重组体转化E.coli BL21大肠杆菌表达宿主菌,酶切鉴定阳性菌落后,分别在M9和LB培养基中用异丙基β-D硫代半乳糖（IPTG,Isopropyl-beta-D-thiogalactoside）诱导表达,SDS-PAGE电泳检测表达的PEDF蛋白,美国I mage-Pro Plus分析系统进行蛋白定量分析。结果 LB和M9培养基中均获得相对分子质量约54×10^3的rPEDF融合蛋白。但LB培养基获得的是rPEDF融合蛋白的包涵体,目的蛋白占总蛋白含量为21.046%,M9培养基获得的是可溶性的rPEDF的融合蛋白,目的蛋白占总蛋白含量的12.31%。结论不同种培养基中均有rPEDF融合蛋白的表达。

The Expression of Recombinant Pigment Epithelium Derived Factor in the Different Medium

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.