Skip to main content Accessibility help
×
Home

Display of H5N1 Avian influenza virus haemagglutinin HA1 on Bacillus thuringiensis cell surface and its immunogenicity for mice

  • Liu Mei (a1) (a2), Lu Lin-Jing (a1), Huang Jun-Yan (a2), Zhang Shu-Huan (a1), Bi Ding-Ren (a1) (a2) and Sun Ming (a2)...

Abstract

The S-layer protein CTC surface display system of Bacillus thuringiensis was used to test the possibility of displaying H5N1 Avian influenza virus (AIV) haemagglutinin HA1 on the cell surface of B. thuringiensis. Two recombinant plasmids, pCTC-HA1P and pCSHA1P, were constructed by replacing the central part below the surface anchor sequence slh of S-layer protein gene ctc with part ha1 gene (ha1p). pCTC-HA1P harboured the fusion gene ctc-ha1p and pCSHA1P the fusion gene csa-ctc-ha1p, csa representing the csaAB operon (very important in anchoring S-layer protein on the bacterial cell surface). Two recombinant B. thuringiensis strains were constructed by electrotransferring recombinant plasmids to B. thuringiensis plasmid-free derivative strain BMB171. Strains obtained were CH (bearing pCSHA1P) and BCCH (bearing pCTC-HA1P as well as the csaAB operon-carrying plasmid pMIL-CSA). The vegetative cells of CH and BCCH were used as antigens in haemagglutination (HA) and haemagglutination inhibition (HI) assays. HA assay showed recombinant HA1 proteins successfully displayed on the cell surface of CH and BCCH. HI assay showed that these recombinant HA1 proteins were specific to standard positive HI (haemagglutination inhibition test) serum of subtype H5 AIV. After immunization of mice with vegetative cells, both CH and BCCH elicited a humoral response to HA1 and exhibited immunogenicity as indicated by enzyme-linked immunosorbent assay (ELISA). ELISA also showed that CH exhibited a higher immunogenicity than BCCH. The strategy developed in this study suggests the possibility of generating a heat-stable and oral veterinary vaccine against AIV with the B. thuringiensis S-layer protein CTC surface display system.

Copyright

Corresponding author

*Corresponding author. E-mail: bidingren@mail.hzau.edu.cn

References

Hide All
Guo, XH, Xiong, Z, Zhou, M, Jia, SF and Xu, Y (1991) The construction of multifunctional shuttle vectors of Bacillus subtilisEscherichia coli. Chinese Journal of Biotechnology 7(3): 224229 (in Chinese with English abstract).
Jiang, YP, Zhang, HB, Li, CJ, et al. (2006) Comparison of protection effects of H5 subtype Avian influenza DNA vaccine optimized with HA gene and expressive vector. Journal of Agricultural Biotechnology 14(3):301306 (in Chinese with English abstract).
Jin, Q (2001) Medicine Molecular Virology. Beijing: Science Press, pp. 646648 (in Chinese).
Li, L and Yu, ZN (1999) Transformation and expression properties of a Bacillus thuringiensis plasmid-free derivative strain BMB171. Chinese Journal of Applied and Environment Biology 5(4):395399 (in Chinese with English abstract).
Li, L, Shao, ZZ and Yu, ZN (2000) Transformation of Bacillus thuringiensis recipient BMB171 by electroporation. Microbiology 27(5):331334 (in Chinese with English abstract).
Mesnage, S, Weber-Levy, M, Haustant, M, Mock, M and Fouet, A (1999) Cell surface-exposed tetanus toxin fragment C produced by recombinant Bacillus anthracis protects against tetanus toxin. Infection and Immunity 67(9): 48474850.
Mesnage, S, Fontaine, T, Mignot, T, Delepierre, M, Mock, M and Fouet, A (2000) Bacterial SLH domain proteins are non-covalently anchored to the cell surface via a conserved mechanism involving wall polysaccharide pyruvylation. EMBO Journal 19(17): 44734484.
Sambrook, J, Fritsch, EF and Maniatis, T (1989) Molecular Cloning: A Laboratory Manual, 2nd ed.New York: Cold Spring Harbor Laboratory Press.
Sára, M and Sleytr, UB (2000) S-layer proteins. Journal of Bacteriology 182(4): 859868.
Schnepf, E, Crickmore, N, van Rie, J, et al. (1998) Bacillus thuringiensis and its pesticidal crystal proteins. Microbiology and Molecular Biology Reviews 62(3): 775806.
Sleytr, UB and Beveridge, TJ (1999) Bacterial S-layers. Trends in Microbiology 7(6): 253260.
Sneath, PHA, Mair, NS, Sharpe, ME and Holt, JG (1986) Bergey's Manual of Systematic Bacteriology. Vol. 2. Baltimore: Williams and Wilkins, pp. 11051139.
Sun, M, Zhu, CG and Yu, ZN (2001) Cloning of parasporal body protein gene resembling S-layer protein genes from Bacillus thuringiensis CTC strain. Acta Microbiologica Sinica 41(2):141147 (in Chinese with English abstract).
Wang, L, Sun, M and Yu, ZN (2004) Capacity of Bacillus thuringiensis S-layer protein displaying polyhistidine peptides on the cell surface. Applied Biochemistry and Biotechnology 119(2): 133143.
Wu, HG, Liu, X, Sun, M and Yu, ZN (2004) Cell surface display of α-amylase and metallothionein by using S-layer protein of Bacillus thuringiensis strain CTC. Acta Microbiologica Sinica 44(5): 658662 (in Chinese with English abstract).
Zhu, LP and Chen, XQ (2000) Common Experimental Methods for Immunology. Beijing: The People's Military Medicine Press, pp. 352356 (in Chinese).

Keywords

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed