RESUMEN
Objective To display of heterologous proteins on the surface of E. coli . Methods The 1653 bp luciferase report gene was knocked in Ipp and ompA genes of chromosome by Red recombine system and selection-counter selection kan/sacB. Results The quantitative analysis results of exogenous lu-ciferase 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-effi-ciently 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.
RESUMEN
To achieve efficient and stable expression of heterologous exogenetic protein or antigen in E. coli chromosome, the luciferase report gene was knocked in lacZ site of chromosome lac operon by using Red recombination system and selection-counterselection kan/sacB technology. The quantitative analysis of exogenous gene expression indicated that the target gene could be efficiently expressed at lacZ site of lac operon. The results confirmed the efficient screening and stable expression of heterologous protein or antigen on chromosome by using the recombinant engineering technique. This study demonstrated that the chromosome could be used as a vector for heterologous protein or antigen and the stable expression of exogenous gene on E. coli chromosome had no side effect on the bacterial growth and propagation.