基于荧光强度快速判断蛋白可溶性方法的建立

摘要

Objective: To construct a technology detecting protein solubility by observing fluorescence. Methods: We selected radiation protective protein CBL502-AA' and it's mutation CBL502-△AA' as objective proteins, EGFP as reporter protein, constructed EGFP-AA' and EGFP-AAA' fusion protein expression vector respectively. The protein solubility was compared by parallel SDS-PAGE and fluorescence observing. Results: The protein solubility detected by these two methods was identical. Conclusion: The fluorescence based protein solubility fast detection method is preliminary found.%目的:建立一种通过观察荧光强度,快速、直观地判断蛋白可溶性的方法.方法:选择抗辐射蛋白CBL502-AA及其突变体CBLS02-△AA'作为目的蛋白,增强型绿色荧光蛋白作为报告分子,分别构建融合蛋白表达载体；通过SDS-PAGE和荧光观察两种手段检测和比较融合蛋白的可溶性.结果:荧光观察融合蛋白的可洛性与SDS-PAGE检测结果一致.结论:建立了一种基于荧光强度,快速、简单、直观的比较蛋白可溶性的方法.