Pallesen L, Poulsen L K, Christiansen G, Klemm P
Department of Microbiology, Technical University of Denmark, Lyngby, Denmark.
Microbiology (Reading). 1995 Nov;141 ( Pt 11):2839-48. doi: 10.1099/13500872-141-11-2839.
The FimH adhesin of type 1 fimbriae has been tested as a display system for heterologous protein segments on the surface of Escherichia coli. This was carried out by introduction of restriction site handles (BglII sites) in two different positions in the fimH gene, followed by in-frame insertion of heterologous DNA segments encoding two reporter sequences. In the selected positions such insertions did not significantly alter the function of the FimH protein with regard to surface location and adhesive ability. The system seemed to be quite flexible, since chimeric versions of the FimH adhesin containing as many as 56 foreign amino acids were transported to the bacterial surface as components of the fimbrial organelles. Furthermore, the foreign protein segments were recognized by insert-specific antibodies when expressed within chimeric proteins on the surface of the bacteria. The results from this feasibility study point to the possibility of using the FimH adhesin as a general surface display system for sizeable protein segments.
1型菌毛的FimH黏附素已被测试作为大肠杆菌表面异源蛋白片段的展示系统。这是通过在fimH基因的两个不同位置引入限制性酶切位点(BglII位点),随后框内插入编码两个报告序列的异源DNA片段来实现的。在选定的位置,这种插入在FimH蛋白的表面定位和黏附能力方面没有显著改变其功能。该系统似乎相当灵活,因为含有多达56个外源氨基酸的FimH黏附素嵌合体版本作为菌毛细胞器的组成部分被转运到细菌表面。此外,当外源蛋白片段在细菌表面的嵌合蛋白中表达时,能被插入特异性抗体识别。这项可行性研究的结果表明,有可能将FimH黏附素用作相当大的蛋白片段的通用表面展示系统。