Wild Strain of duck pathogenic Escherichia coli was isolated from avian pathogenic colibacillosis suspected duck in a waterfowl farm in Jiangsu province. The isolated strain was subject to serotype identification, cluster analysis, virulence gene detection, E protein expression and bacterial ghosts preparing, and morphology observation. After inactivation and sterility test, ducklings were immunized with bacterial ghost and serum IgG levels were detected. The results showed that the serotype of the strain is O24 and the strain belongs to group B1. It has virulence genes fimC, csgA and iroN. The lysis rate of duck pathogenic Escherichia coli was 99.96%. The results of transmission electron microscopy showed that there were obvious channels on the surface of bacterial ghosts prepared. We found that the cytoplasm overflowed from the channels and the cell membrane was shrank and deformed. The results of antibody level test showed that the serum IgG level of ducklings in bacterial ghost group increased significantly after secondary immunization. It can be seen that by transforming the recombinant plasmid pBV221-E into duck pathogenic Escherichia coli isolate and regulating the temperature of bacterial culture to induce the expression of E protein. The bacterial ghost of wild strain O24 of B1 group of duck pathogenic Escherichia coli were constructed in this study and it owns capacity to induce humoral immunity.