Indian Journal Of Animal Research

Cloning and Expression of Fasciola gigantica Cathepsin-B Recombinant Proteins

DOI: 10.18805/ijar.B-3959    | Article Id: B-3959 | Page : 333-339
Citation :- Cloning and Expression of Fasciola gigantica Cathepsin-B Recombinant Proteins.Indian Journal Of Animal Research.2021.(55):333-339
Andleeb Aftab, Savita Bisen, Rohit Lall, Shobha Yadav, Silamparasan, M, O.K. Raina rainaok@rediffmail.com
Address : Divion of Parasitology,  Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India. 
Submitted Date : 30-11-2019
Accepted Date : 17-03-2020
First Online:

Abstract

Fasciola gigantica cathepsin (cysteine) proteases are potential diagnostic antigens for animal and human fasciolosis. These include cathepsin-L proteases that have been exploited in the diagnosis of animal fasciolosis. However, no scientific data on the diagnostic potential of F. gigantica cathepsin B proteases is available. Therefore, three recombinant antigens of F. gigantica viz. cathepsin (cat) B-1, cat B-2 and cat B-3 were expressed in prokaryotic expression system. The recombinant antigens were purified under denaturing conditions by Nickel affinity chromatography and an optimal level of the recombinant proteins was obtained. These recombinant proteins will further be evaluated for their potential in the early prepatent diagnosis of F. gigantica infection in domestic ruminants.

Keywords

Cathepsin B Fasciola gigantica Recombinant antigen Serodiagnosis

References

  1. Bardhan, D., Kumar, R.R., Nigam, S., Mishra, H. and Bhoj, S. (2014). Estimation of milk losses due to fasciolosis in Uttarakhand. Agricultural Economics Research Review. 27: 281-288. 
  2. Berasain, P., Goni, F., McGonigle, S., Dowd, A.J., Dalton, J.P., Frangione, B. and Carmona, C. (1997). Proteinases secreted by Fasciola hepatica degrade extra cellular matrix and basement membrane components. Journal of Parasitology. 83: 1-5.
  3. Cancela, M., Acosta, D., Rinaldi, G., Silva, E., Dura´n, R., Roche, L., Zaha, A., Carmona, C. and F. Tort, J. (2008). A distinctive repertoire of cathepsins is expressed by juvenile invasive Fasciola hepatica. Biochimie. 90: 1461-1475.
  4. Chantree, P., Wanichanon, C., Phatsara, M., Meemon, K. and Sobhon, P. (2012). Characterization and expression of cathepsin B2 in Fasciola gigantica. Experimental Parasitology. 132: 249-256.
  5. Charlier, J., Duchateau, L., Claerebout, E., Williams, D. and Vercruysse, J. (2007). Associations between anti-Fasciola hepatica antibody levels in bulk-tank milk samples and production parameters in dairy herds. Preventive Veterinary Medicine. 78: 57-66. 
  6. Chick, B.F., Loverdale, O.R. and Jackson, A.R.B. (1980). Production effects of liver fluke infection in beef cattle. Australian Veterinary Journal. 20: 123-127.
  7. Dalton, J.P., Neil, S.O., Stack, C., Collins, P., Walshe, A., Sekiya, M., Doyle, S., Mulcahy, G., Hoyle, D., Khaznadji, E., Moire, N., Brennan, G., Mousley, A., Kreshchenko, N., Maule A. G and Donnelly, S.M. (2003). Fasciola hepatica cathepsin L-like proteases: biology, function and potential in the development of ûrst generation liver ûuke vaccines. International Journal for Parasitology. 33: 1173-1181.
  8. Dixit, A.K., Dixit, P. and Sharma, R.L. (2003). Immunodiagnostic / protective role of cathepsin L cysteine proteinases secreted by Fasciola species. Veterinary Parasitology. 154: 177-184.
  9. Dixit, A.K., Yadav, S.C. and Sharma, R.L. (2002). The 28 kDa Fasciola gigantica cysteine proteinase in the diagnosis of prepatent ovine fasciolosis. Veterinary Parasitology. 109: 233-247.
  10. Dowd, A.J., Smith, A.M., McGonigle, S. and Dalton, J.P. (1994). Purification and characterization of a second cathepsin-    L proteinase secreted by the parasitic trematode Fasciola hepatica. European Journal of Biochemistry. 223: 91-98.
  11. Graham-Brown, J., Williams D. J.L., Skuce, P., Zadoks, R.N., Dawes, S., Swales, H., Van-Dijk, J. (2019). Composite Fasciola hepatica faecal egg sedimentation test for cattle. Veterinary Record. 184: 589. 
  12. Kleiman, F., Pietrokovsky, S., Gil S., Wisnivesky-Colli, C. (2005). Comparison of two coprological methods for the veterinary diagnosis of fasciolosis. Arq. Bras. Med. Vet. Zootec. 57 (2). Belo Horizonte Print version ISSN 0102-0935 On-line version ISSN 1678-4162.
  13. Laemmli, U.K (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680-685.
  14. Law, R.H., Smooker, P.M., Irving J.A., Piedrafita, D., Ponting, R., Kennedy, N.J, Whisstock, J.C, Pike, R.N, Spithill, T.W. (2003). Cloning and expression of the major secreted cathepsin B-like protein from juvenile Fasciola hepatica and analysis of immunogenicity following liver fluke infection. Infection and Immunity. 71: 6921-6932.
  15. Meemon, K., Grams, R., Vichasri-Grams, S., Hofmann, A., Korge, G., Viyanant, V., Upatham, E. S., Habe, S. and Sobhon, P. (2004). Molecular cloning and analysis of stage and tissue-specific expression of cathepsin B encoding genes from Fasciola gigantica. Molecular and Biochemical Parasitology. 136: 1-10.
  16. Mehra, U.R., Verma, A.K., Dass, R.S., Sharma, R.L. and Yadav, S.C. (1999). Effects of Fasciola gigantica infection on growth and nutrient utilization of buffalo calves. Veterinary Research. 145: 699-702. 
  17. Mezo, M., Gonzalez-Warleta, M., Castro-Hermida, J.A., Muino, L. and Ubeira, F.M. (2011). Association between anti-F. hepatica antibody levels in milk and production losses in dairy cows. Veterinary Parasitology. 180: 237-242.
  18. Nyirenda, S.S., Sakala, M., Moonde, L., Kayesa, E., Fandamu, P., Banda, F. and Sinkala, Y (2019). Prevalence of bovine
  19. fascioliasis and economic impact associated with liver condemnation in abattoirs in Mongu district of Zambia. BMC Veterinary Research. 15: 33.
  20. Raina, O.K., Yadav, S.C., Sriveny, D. And Gupta, S.C (2006). Immunodiagnosis of bubaline fasciolosis with Fasciola gigantica cathepsin-L and recombinant cathepsin L 1-D proteases. Acta Tropica. 98:145-151.
  21. Sambrook, J.F and Russell, D.W. (2001). Molecular Cloning: A Laboratory Manual, 3rd
  22. Sethadavit, M., Meemon, K., Jardim, A., Spithill, T.W. and Sobhon, P. (2009). Identification, expression and immunolocalization of cathepsin B3, a stage-specific antigen expressed by juvenile Fasciola gigantica. Acta Tropica. 112: 164-173.
  23. Sriveny, D., Raina, O.K., Yadav, S.C., Chandra, D., Jayrawa, A.K., Singh, M., Velusamy, R. and Singh, B.P. (2006). Cathepsin L cysteine proteinase in the diagnosis of bovine Fasciola gigantica infection. Veterinary Parasitology. 135: 25-31.
  24. Towbin, H., Staehelin, T. and Gordon, J. (1979). Electrophoretic transfer of proteins from acrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences. 76: 4350-4354.
  25. Varghese, A., Raina, O.K., Nagar, G., Garg, R., Banerjee, P.S., Maharana, B. R. and Kollannur, J.D. (2012). Development of cathepsin-L cysteine proteinase based dot-enzyme-linked immunosorbent assay for the diagnosis of Fasciola gigantica infection in buffaloes. Veterinary Parasitology. 183: 382-385.
  26. Yadav, S.C., Saini, M., Raina, O.K., Nambi, P.A., Jadav, K. and Sriveny, D. (2005). Fasciola gigantica cathepsin-L cysteine proteinase in the detection of early experimental fasciolosis in ruminants. Parasitology Research. 97: 527-534.

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