Abstract：
To enable an efficient bacterial cell surface display with effective protein expression and cell surface loading ability via autotransporter for potential vaccine development applications, the inner membrane protein translocation efficiency was investigated via a trial-and-error strategy by replacing the original unusual long signal peptide of E. coli Ag43 with 11 different signal peptides. The receptor-binding domain (RBD) of coronavirus was used as a neutral display substrate to optimize the expression conditions, and the results showed that signal peptides from PelB, OmpC, OmpF, and PhoA protein enhance the bacterial cell surface display efficiency of RBD. In addition, the temperature has also a significant effect on the autodisplay efficiency of RBD. Our data provide further technical basis for the biotechnological application of Ag43 as a bacterial surface display carrier system and further potential application in vaccine development.
摘要：
为了使有效的细菌细胞表面展示具有有效的蛋白质表达和细胞表面负载能力，通过自身转运蛋白用于潜在的疫苗开发应用。本研究通过试错策略,用11种不同的信号肽替换大肠杆菌Ag43的原始异常长信号肽,研究了内膜蛋白易位效率.以冠状病毒的受体结合域(RBD)作为中性展示底物,优化表达条件,结果显示来自PelB的信号肽，OmpC,OmpF,和PhoA蛋白增强RBD的细菌细胞表面展示效率。此外,温度对RBD的自动显示效率也有显著影响。我们的数据为Ag43作为细菌表面展示载体系统的生物技术应用以及在疫苗开发中的进一步潜在应用提供了进一步的技术基础。
