Abstract
Fragment E of ureB (ureBE) was cloned from a clinical isolate of Helicobacter pylori. A prokaryotic expression vector, pAMJ399, with the ureB fragment E and the Staphylococcus aureus protein A anchor fragment (spaX), was constructed. The fusion protein was expressed under the control of the P170 promoter in Lactococcus lactis. Western blot assay of lactococcal cell wall extracts with a polyclonal chicken antiserum confirmed the immunity of the expressed recombinant protein which was located on the cell surface. These results provide the first report of a surface display system in lactic acid bacteria for the delivery of oral vaccines against Helicobacter pylori.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akada JK, Shirai M, Takeuchi H, Tsuda M, Nakazawa T (2000) Identification of the urease operon in Helicobacter pylori and its control by mRNA decay in response to pH. Mol Microbiol 36:1071–1084
Challacombe SJ, Rahman D, Jeffery H, Davis SS, O’Hagan DT (1992) Enhanced secretory IgA and systemic IgG antibody responses after oral immunization with biodegradable microparticles containing antigen. Immunology 76:164–168
Corthesy B, Boris S, Isler P, Grangette C, Mercenier A (2005) Oral immunization of mice with lactic acid bacteria producing Helicobacter pylori urease B subunit partially protects against challenge with Helicobacter felis. J Infect Dis 192:1441–1449
Corthesy-Theulaz I, Porta N, Glauser M, Sarage E, Vaney AC, Haas R, Kraehenbuhl JP, Blum AL, Michetti P (1995) Oral immunization with Helicobacter pylori urease B subunit as a treatment against Helicobacteri infection in mice. Gastroenterology 109:115–121
Forman D, Newell DG, Fullerton F, Yarnell JW, Stacey AR, Wald N, Sitas F (1991) Association between infection with Helicobacter pylori and risk of gastric cancer: evidence from a prospective investigation. Br Med J 302:1302–1305
Gu Q, Yao L, Li J, Zhu MY (2006) Expression of the C-terminal 26 kDa fragment of Helicobacter pylori UreB in Escherichia coli and identification of the recombinant protein immunity. J Zhejiang Univ 32:387–390
Holo H, Nes IF (1989) High-frequency transformation, by electroporation, of Lactococcus lactis subsp. cremoris grown with glycine in osmotically stabilized media. Appl Environ Microbiol 55:3119–3123
Kleanthous H, Myers GA, Georgakopoulos KM, Tibbitts TJ, Ingrassia JW, Gray L, Ding R, Zhang ZZ, Lei W, Nichols R, Lee CK, Ermak TH, Monath TP (1998) Rectal and intranasal immunizations with recombinant urease induce distinct local and serum immune responses in mice and protect against Helicobacter pylori infection. Infect Immun 66:2879–2886
Klijn N, Weerkamp AH, de Vos WM (1995) Genetic marking of Lactococcus lactis shows its survival in the human gastrointestinal tract. Appl Environ Microbiol 61:2771–2774
Labigne A, Cussac V, Courcoux P (1991) Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity. J Bacteriol 173:1920–1931
Lee MH, Roussel Y, Wilks M, Tabaqchali S (2001) Expression of Helicobacter pylori urease subunit B gene in Lactococcus lactis MG1363 and its use as a vaccine delivery system against H. pylori infection in mice. Vaccine 19:3927–3935
Marshall BJ (1994) Helicobacter pylori. Am J Gastroenterol 89:S116–S118
Michetti P (1997) Vaccine against Helicobacter pylori: fact or fiction? Gut 41:728–730
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York, pp 1.21–1.52, 2.60–2.80, 7.3–7.35, 9.14, 9.22
Schneewind O, Mihaylova-Petkov D, Model P (1993) Cell wall sorting signals in surface proteins of gram-positive bacteria. EMBO J 12:4803–4811
Seegers JF (2002) Lactobacilli as live vaccine delivery vectors: progress and prospects. Trends Biotechnol 20:508–515
Shuttleworth HL, Duggleby CJ, Jones SA, Atkinson T, Minton NP (1987) Nucleotide sequence analysis of the gene for protein A from Staphylococcus aureus Cowan 1 (NCTC 8530) and its enhanced expression in Escherichia coli. Gene 58:283–295
Steidler L, Viaene J, Fiers W, Remaut E (1998) Functional display of a heterologous protein on the surface of Lactococcus lactis by means of the cell wall anchor of Staphylococcus aureus protein A. Appl Environ Microbiol 64:342–345
Tomb JF, White O, Kerlavage AR et al (1997) The complete genome sequence of the gastric pathogen Helicobacter pylori. Nature 388:539–547
Wells JM, Robinson K, Chamberlain LM, Schofield KM, Le Page RWF (1996) Lactic acid bacteria as vaccine delivery vehicles. Antonie Leeuwenhoek 70:317–330
Acknowledgments
This work was supported by the National Natural Science Foundation of China (No. 30300017), the Natural Science Foundation of Zhejiang Province (No. R306425), and the program for science and technology from Zhejiang Province (Nos. 2005C21043; 2007C12037).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Song, D., Gu, Q. Surface expression of Helicobacter pylori urease subunit B gene E fragment on Lactococcus lactis by means of the cell wall anchor of Staphylococcus aureus protein A. Biotechnol Lett 31, 985–989 (2009). https://doi.org/10.1007/s10529-009-9965-9
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-009-9965-9