Skip to main content
Log in

Gene Cloning, Expression, and Characterization of Recombinant Aerolysin from Aeromonas hydrophila

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Aerolysin is a significant virulent toxin protein secreted by Aeromonas hydrophila; it produces deep wound infections and hemorrhagic septicemia. The complete aerolysin gene (1,482 bp) was amplified from A. hydrophila. Furthermore, it was cloned and expressed into Escherichia coli BL21(DE3) codon plus RP cells using 0.5 mM IPTG for induction. The protein size was 54 kDa as estimated by SDS-PAGE, and it was purified by Ni–NTA affinity chromatography. Anti-His antibodies were used to characterize the expressed aerolysin by Western blotting and showed hemolytic activity with fish red blood cells. Aerolysin may be used as immunoassays for earlier control of A. hydrophila and is also compatible for vaccination.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Abrami, L., Fivaz, M., Glauser, P.-E., Parton, R. G., & van der Goot, F. G. (1998). Journal of Cell Biology, 140, 525–540.

    Article  CAS  Google Scholar 

  2. Wang, A., Wada, A., Yahiro, K., Nomura, T., Fujii, Y., Okamato, K., et al. (1999). Microbial Pathogenesis, 27, 215–221.

    Article  CAS  Google Scholar 

  3. Howard, S. P., Garlnd, W. J., Green, J. M., & Buckley, J. T. (1987). Journal of Bacteriology, 169, 2869–2871.

    CAS  Google Scholar 

  4. Parker, M. W., Buckley, J. T., Postma, J. P. M., Tucker, A. D., Leonard, K., Pattus, F., et al. (1994). Nature, 367, 292–295.

    Article  CAS  Google Scholar 

  5. Chakraborty, T., Huhle, B., Hof, H., Bergbauer, H., & Goebel, W. (1987). Infection and Immunity, 55, 2274–2280.

    CAS  Google Scholar 

  6. Heuzenroeder, M. W., Wong, C. Y., & Flower, R. L. (1999). FEMS microbiology Letters, 174, 131–136.

    Article  CAS  Google Scholar 

  7. Khan, A. A., Kim, E., & Cerniglia, C. E. (1998). Applied Environmental Microbiology, 64(7), 2473–8.

    CAS  Google Scholar 

  8. Wong, C. Y., Heuzenroeder, M. W., & Flower, R. L. (1998). Microbiology, 144(2), 291–8.

    Article  CAS  Google Scholar 

  9. Khushiramani, R., Girisha, S. K., Karunasagar, I., & Karunasagar, I. (2007). Protein Expression and Purification, 51(2), 303–7.

    Article  CAS  Google Scholar 

  10. Singh, V., & Somvanshi, P. (2009). Letters in Drug Design and Discovery, 6(3), 215–223.

    Article  CAS  Google Scholar 

  11. Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.

    Google Scholar 

  12. Zhu, D., Li, A., Wang, J., Li, M., & Cai, T. (2007). Indian Journal of Biochemistry and Biophysics, 44(4), 204–208.

    CAS  Google Scholar 

  13. Austin, B., & Austin, D. A. (1999). Bacterial fish pathogens: Diseases in farmed and wild fish. Chichester, UK: Praxis Publishing.

    Google Scholar 

  14. Janda, J. M., Abbott, S. L., Cheung, W. K., & Hanson, D. F. (1994). Journal of Clinical Microbiology, 32(8), 1850–1854.

    CAS  Google Scholar 

  15. Merino, S., Rubires, X., Knochel, S., & Tomas, J. M. (1995). International Journal of Food Microbiology, 28, 157–168.

    Article  CAS  Google Scholar 

  16. Pemberton, J. M., Kidd, S. P., & Schmidt, R. (1997). FEMS Microbiology Letters, 152, 1–10.

    Article  CAS  Google Scholar 

  17. Chopra, A. K., Peterson, C. W., & Jin, G. F. (1993). Canadian Journal of Microbiology, 39, 513–523.

    Article  CAS  Google Scholar 

  18. Howard, S. P., & Buckley, J. T. (1985). Journal of Bacteriology, 163, 336–340.

    CAS  Google Scholar 

  19. Singh, V., Rathore, G., Kapoor, D., Mishra, B. N., & Lakra, W. S. (2008). Indian Journal of Microbiology, 48(4), 453–458.

    Article  CAS  Google Scholar 

  20. Singh, V., Rathore, G., Kumar, G., Swaminathan, T. R., Sood, N., Kapoor, D., et al. (2007). Indian Veterinary Journal, 84, 900–02.

    Google Scholar 

  21. Singh, V., Somvanshi, P., Rathore, G., Kapoor, D., & Mishra, B. N. (2008). Protein Expression and Purification, 65(1), 1–7.

    Article  CAS  Google Scholar 

Download references

Acknowledgement

The authors are grateful to Dr. S. Ayyappan, DDG (Fy) Indian Council of Agricultural Research, New Delhi. The authors are thankful the Dr. W.S. Lakra, Director, National Bureau of Fish Genetic Resources, Lucknow, providing the timely suggestion and valuable guidance.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Vijai Singh.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Singh, V., Somvanshi, P., Rathore, G. et al. Gene Cloning, Expression, and Characterization of Recombinant Aerolysin from Aeromonas hydrophila . Appl Biochem Biotechnol 160, 1985–1991 (2010). https://doi.org/10.1007/s12010-009-8752-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-009-8752-3

Keywords

Navigation