ZFIN ID: ZDB-PUB-110519-22
Iron-regulated lysis of recombinant Escherichia coli in host to release protective antigen and confer biological containment
Guan, L., Mu, W., Champeimont, J., Wang, Q., Wu, H., Xiao, J., Lubitz, W., Zhang, Y., and Liu, Q.
Date: 2011
Source: Infection and Immunity   79(7): 2608-18 (Journal)
Registered Authors: Liu, Qin
Keywords: none
MeSH Terms:
  • Aeromonas hydrophila/immunology
  • Animals
  • Antigens, Bacterial/genetics
  • Antigens, Bacterial/immunology*
  • Bacterial Vaccines*/administration & dosage
  • Bacterial Vaccines*/genetics
  • Bacterial Vaccines*/immunology
  • Bacteriolysis
  • Escherichia coli/genetics*
  • Escherichia coli/immunology*
  • Escherichia coli Proteins/genetics
  • Escherichia coli Proteins/metabolism
  • Flatfishes/immunology
  • Glyceraldehyde-3-Phosphate Dehydrogenases/genetics
  • Glyceraldehyde-3-Phosphate Dehydrogenases/immunology*
  • Gram-Negative Bacterial Infections/immunology
  • Green Fluorescent Proteins/biosynthesis
  • Green Fluorescent Proteins/genetics
  • Iron/metabolism*
  • Isopropyl Thiogalactoside/metabolism
  • Plasmids
  • Promoter Regions, Genetic
  • Signal Transduction
  • Vaccines, Synthetic/administration & dosage
  • Vaccines, Synthetic/genetics
  • Vaccines, Synthetic/immunology
  • Zebrafish/immunology
PubMed: 21536797 Full text @ Infect. Immun.

Recombinant bacterial vector vaccine is an attractive vaccination strategy to induce immune response to a carried protective antigen. The superiorities of live bacterial vectors include mimicry of a natural infection, intrinsic adjuvant properties and the potential for administration by mucosal routes. Escherichia coli is a simple and efficient vector system for production of exogenous proteins. In addition, many strains are nonpathogenic and avirulent making it a good candidate for use in recombinant vaccine design. In this study, we screened 23 different iron-regulated promoters in E. coli BL21(DE3) vector and found one, PviuB, with characteristics suitable for our use. We fused PviuB with lysis gene E, establishing an in vivo inducible lysis circuit. The resulting in vivo lysis circuit was introduced into a strain also carrying an IPTG-inducible PT7-controlled protein synthesis circuit, forming a novel E. coli-based protein delivery system. The recombinant E. coli produced a large amount of antigen in vitro, and could deliver the antigen into zebrafish after vaccination via injection. The strain subsequently lysed in response to the iron-limiting signal in vivo, implementing antigen release and biological containment. The gapA gene, encoding the protective antigen GAPDH from the fish pathogen Aeromonas hydrophila LSA34, was introduced into the E. coli-based protein delivery system, and the resultant recombinant vector vaccine was evaluated in turbot (Scophtalmus maximus). Over 80% of the vaccinated fish survived challenge with A. hydrophila LSA34, suggesting that the E. coli-based antigen delivery system had great potential in bacterial vector vaccine application.