Abstract: 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.