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目的 优化重组大肠杆菌热不稳定肠毒素B亚单位 (rLTB)工程菌 (VSP6 0 )高效表达的条件。方法 以UVP凝胶照相系统扫描以及改良Lowry′s法检测rLTB表达量并分析各种因素对其影响。利用SephacrylS 10 0凝胶层析纯化rLTB蛋白。结果 工程菌 30℃振荡培养至吸光度 (A6 0 0 )值达0 .2～ 0 .3时加IPTG(终浓度为 0 .5mmol L) ,诱导培养 18～ 2 2h为rLTB蛋白表达的最佳条件。SephacrylS 10 0凝胶柱一步层析后 ,rLTB蛋白纯度可达 98.1%。结论 本研究所用的培养和纯化工艺简单易行 ,可获得高产量、高纯度的rLTB蛋白。
Objective To optimize the conditions for efficient expression of the recombinant E. coli heat labile enterotoxin subunit B (rLTB) engineered bacteria (VSP60). Methods UVP gel imaging system scanning and modified Lowry’s method to detect rLTB expression and analysis of various factors on its impact. The rLTB protein was purified by Sephacryl S10 gel filtration. Results The optimal conditions for rLTB protein expression were induced by IPTG (final concentration was 0.5 mmol L) after incubation at 30 ℃ with shaking until absorbance (A6 0 0) reached 0.2-2.3. . After one-step chromatography on Sephacryl S 10 0 gel column, the purity of rLTB protein reached 98.1%. Conclusion The culture and purification techniques used in this study were simple and easy to obtain high yield and high purity rLTB protein.