应用Red重组工程技术在大肠杆菌外膜展示表达外源蛋白

Display of heterologous proteins on the surface of E. coli by Red recombineering

目的将外源蛋白或抗原稳定表达展示于大肠杆菌的外膜上。方法应用大肠杆菌DY330介导的重组工程系统和kan/sacB无痕迹修饰技术，将长度为1653 bp的荧光素酶报告基因（luc）敲入DY330染色体与外膜蛋白基因lpp、ompA融合。结果报告基因定量分析结果证明：外源荧光素酶（1uciferase，Luc）能够与外膜蛋白融合表达于细菌外膜，Lpp-OmpA-Luc融合蛋白表达于细菌外膜的效率最高。结论外源蛋白能够稳定表达于细菌外膜，不会影响细菌的生长繁殖。

Objective To display of heterologous proteins on the surface of E. coli. Methods The 1653 bp luciferase report gene was knocked in lpp and ompA genes of chromosome by Red recombine system and selection-counter selection kan/sacB. Results The quantitative analysis results of exogenous luciferase expression displayed that it could be expressed as fusion with the outer membrane proteins on the cell surface. The fusion protein of foreign protein and outer membrane protein Lpp-OmpA-Luc could be high-efficiently displayed on cell surface. Conclusion The stable expression of exogenous gene on the surface of E. coli had no effect on the bacterial growth and propagation.