大肠杆菌表面展示结核分枝杆菌噬菌体结合肽

Display of Mycobacterium Tuberculosis Binding Peptide on Escherichia Coli Cell Surface

构建重组表达载体,将分枝杆菌噬菌体中分离出小分子肽段PK34的基因与大肠杆菌外膜蛋白OmpC基因相融合,并利用OmpC自身启动子将PK34及组氨酸标签在大肠杆菌外膜上进行重组蛋白表达,借助磁珠建立表面展示体系.结果表明,利用外膜蛋白OmpC的穿膜特性,能够在大肠杆菌表面对分枝杆菌噬菌体PK34肽段进行展示.通过表面展示的PK34与分枝杆菌进行有效结合,从样品中对分枝杆菌进行分离.为今后临床样本中结核分枝杆菌的分离与筛选方法提供新思路.

A recombinant expression vector was constructed to fuse the gene of small molecule peptide PK34 isolated from mycobacteria phages with the E.coli outer membrane protein OmpC gene,express recombinant PK34 and histidine labels on the outer membrane using the OmpC own promoter,and establish a surface display system with magnetic beads,and the surface display system was establisted with magnetic beads.The results showed that the peptide PK34 from mycobacteriophage could be displayed on the surface of E.coli by adopting the characteristic of outer membrane protein OmpC.Mycobacterium could be isolated through effective binding of PK34 within the surface display system.This study provides a new insight of the isolation and screening of Mycobacterium from clinical trial in the future.