To synthesize and secrete heterologous proteins in an attenuated Vibrio anguillarum strain for potential multivalent live vaccine development, different antigen-delivery systems based on bacterial-originated secretion signal peptides (SPs) were designed and identified in this work. Four SPs were derived from hemolysin of Escherichia coli, RTX protein of V. cholerae, hemolysin of V. anguillarum, zinc-metalloprotease of V. anguillarum, respectively, and their abilities to support secretion of green fluorescent protein (GFP) in an attenuated V. anguillarum strain MVAV6203 were assayed. Immunodetection of GFP showed that the capability of the tested signal leaders to direct secretion of GFP varied greatly. Although all the four signal peptide-fused GFPs could be expressed correctly and trapped intracellularly in recombinant strains, only the EmpA signal peptide could confer efficient secretion to GFP. For the investigation of its potential application in live bacteria carrier vaccines, a heterologous protein EseB of Edwardsiella tarda was fused to the SPempA antigen-delivery system and introduced into the strain MVAV6203. Further analysis of EseB demonstrated that the constructed SPempA antigen-delivery system could be used to secrete foreign protein in attenuated V. anguillarum and be available for carrier vaccines development.
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We thank Dr. Biao Kan (The Chinese Academy of Preventive Medicine) and Dr. Andersen (Technical University of Denmark) for the kindness in sending strains and plasmids used in this study. This work was supported by the National Natural Science Foundation of China (30600460), the National High Technology Research and Development Program of China (2006AA100310), and the State Key Development Program of Basic Research of China (2006CB101800).
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Zhou, L., Liu, Q., Wang, Q. et al. Secretory delivery of heterologous proteins in attenuated Vibrio anguillarum for potential use in vaccine design. Appl Microbiol Biotechnol 79, 1027–1034 (2008). https://doi.org/10.1007/s00253-008-1511-5
- Green fluorescent protein
- Signal peptide
- Vibrio anguillarum