Hostname: page-component-cd9895bd7-fscjk Total loading time: 0 Render date: 2024-12-27T10:49:33.219Z Has data issue: false hasContentIssue false

Paramyosin from the parasitic mite Sarcoptes scabiei: cDNA cloning and heterologous expression

Published online by Cambridge University Press:  07 August 2001

J. G. MATTSSON
Affiliation:
Department of Parasitology (SWEPAR), National Veterinary Institute, SE-751 89 Uppsala, Sweden
E. L. LJUNGGREN
Affiliation:
Department of Parasitology (SWEPAR), National Veterinary Institute, SE-751 89 Uppsala, Sweden
K. BERGSTRÖM
Affiliation:
Department of Parasitology (SWEPAR), National Veterinary Institute, SE-751 89 Uppsala, Sweden

Abstract

The burrowing mite Sarcoptes scabiei is the causative agent of the highly contagious disease sarcoptic mange or scabies. So far, there is no in vitro propagation system for S. scabiei available, and mites used for various purposes must be isolated from infected hosts. Lack of parasite-derived material has limited the possibilities to study several aspects of scabies, including pathogenesis and immunity. It has also hampered the development of high performance serological assays. We have now constructed an S. scabiei cDNA expression library with mRNA purified from mites isolated from red foxes. Immunoscreening of the library enabled us to clone a full-length cDNA coding for a 102.5 kDa protein. Sequence similarity searches identified the protein as a paramyosin. Recombinant S. scabiei paramyosin expressed in Escherichia coli was recognized by sera from dogs and swine infected with S. scabiei. We also designed a small paramyosin construct of about 17 kDa that included the N-terminal part, an evolutionary variable part of the helical core, and the C-terminal part of the molecule. The miniaturized protein was efficiently expressed in E. coli and was recognized by sera from immunized rabbits. These data demonstrate that the cDNA library can assist in the isolation of important S. scabiei antigens and that recombinant proteins can be useful for the study of scabies.

Type
Research Article
Copyright
2001 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

ARLIAN, L. G. (1989). Biology, host relations, and epidemiology of Sarcoptes scabiei. Annual Review of Entomology 34, 139161.CrossRefGoogle Scholar
ARLIAN, L. G., MORGAN, M. S., VYSZENSKI-MOHER, D. L. & STEMMER, B. L. (1994). Sarcoptes scabiei: the circulating antibody response and induced immunity to scabies. Experimental Parasitology 78, 3750.CrossRefGoogle Scholar
BECKER, K. D., O'DONNELL, P. T., HEITZ, J. M., VITO, M. & BERNSTEIN, S. I. (1992). Analysis of Drosophila paramyosin: identification of a novel isoform which is restricted to a subset of adult muscles. Journal of Cell Biology 116, 669681.CrossRefGoogle Scholar
BORNSTEIN, S., THEBO, P. & ZAKRISSON, G. (1996). Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of canine sarcoptic mange. Veterinary Dermatology 7, 2128.CrossRefGoogle Scholar
BORNSTEIN, S. & WALLGREN, P. (1997). Serodiagnosis of sarcoptic mange in pigs. Veterinary Record 141, 812.Google Scholar
BORNSTEIN, S. & ZAKRISSON, G. (1993). Humoral antibody response to experimental Sarcoptes scabiei var. vulpes infection in the dog. Veterinary Dermatology 4, 107110.CrossRefGoogle Scholar
BURGESS, I. (1994). Sarcoptes scabiei and scabies. Advances in Parasitology 33, 235292.CrossRefGoogle Scholar
CHOSIDOW, O. (2000). Scabies and pediculosis. Lancet 355, 819826.CrossRefGoogle Scholar
DAHMEN, A., GALLIN, M., SCHUMACHER, M. & ERTTMANN, K. D. (1993). Molecular cloning and pre-mRNA maturation of Onchocerca volvulus paramyosin. Molecular and Biochemical Parasitology 57, 335338.CrossRefGoogle Scholar
DEVEREUX, J., HAEBERLI, P. & SMITHIES, O. (1984). A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Research 12, 387395.CrossRefGoogle Scholar
HOLLANDERS, W., VERCRUYSSE, J., RAES, S. & BORNSTEIN, S. (1997). Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of sarcoptic mange in swine. Veterinary Parasitology 69, 117123.CrossRefGoogle Scholar
JACOBS, E. & CLAD, A. (1986). Electroelution of fixed and stained membrane proteins from preparative sodium dodecyl sulfate-polyacrylamide gels into a membrane trap. Analytical Biochemistry 154, 583589.CrossRefGoogle Scholar
KALINNA, B. H. & MCMANUS, D. P. (1997). A vaccine against the Asian schistosome, Schistosoma japonicum: an update on paramyosin as a target of protective immunity. International Journal for Parasitology 27, 12131219.CrossRefGoogle Scholar
LACLETTE, J. P., LANDA, A., ARCOS, L., WILLMS, K., DAVIS, A. E. & SHOEMAKER, C. B. (1991). Paramyosin is the Schistosoma mansoni (Trematoda) homologue of antigen B from Taenia solium (Cestoda). Molecular and Biochemical Parasitology 44, 287295.CrossRefGoogle Scholar
LANDA, A., LACLETTE, J. P., NICHOLSON-WELLER, A. & SHOEMAKER, C. B. (1993). cDNA cloning and recombinant expression of collagen-binding and complement inhibitor activity of Taenia solium paramyosin (AgB). Molecular and Biochemical Parasitology 60, 343347.CrossRefGoogle Scholar
LIMBERGER, R. J. & MCREYNOLDS, L. A. (1990). Filarial paramyosin: cDNA sequences from Dirofilaria immitis and Onchocerca volvulus. Molecular and Biochemical Parasitology 38, 271280.CrossRefGoogle Scholar
MÖRNER, T. (1992). Sarcoptic mange in Swedish wildlife. Revue Scientific et technique Office International des Epizooties 11, 11151121.Google Scholar
NORMAZNAH, Y., SANIAH, K., NAZMA, M., MAK, J. W., KRISHNASAMY, M. & HAKIM, S. L. (1996). Seroprevalence of Sarcoptes scabiei var. canis antibodies among aborigines in peninsular Malaysia. Southeast Asian Journal of Tropical Medicine and Public Health 27, 5356.Google Scholar
PRYOR, K. D. & LEITING, B. (1997). High-level expression of soluble protein in Escherischia coli using a His6-tag and maltose-binding-protein double-affinity fusion system. Protein Expression and Purification 10, 309319.CrossRefGoogle Scholar
RIGGS, P. (1997). Detection and isolation of recombinant protein based on binding affinity reporter. Maltose binding protein. Methods in Molecular Biology 63, 85101.Google Scholar
SCHLESINGER, I., OELRICH, D. M. & TYRING, S. K. (1994). Crusted (Norwegian) scabies in patients with AIDS: the range of clinical presentations. Southern Medical Journal 87, 352356.CrossRefGoogle Scholar
TELLAM, R. L. & BOWLES, V. M. (1997). Control of blowfly strike in sheep: current strategies and future prospects. International Journal for Parasitology 27, 261273.CrossRefGoogle Scholar
WALTON, S. F., CHOY, J. L., BONSON, A., VALLE, A., MCBROOM, J., TAPLIN, D., ARLIAN, L., MATHEWS, J. D., CURRIE, B. & KEMP, D. J. (1999). Genetically distinct dog-derived and human-derived Sarcoptes scabiei in scabies-endemic communities in northern Australia. American Journal of Tropical Medicine and Hygiene 61, 542547.CrossRefGoogle Scholar
WILLADSEN, P. (1997). Novel vaccines for ectoparasites. Veterinary Parasitology 71, 209222.CrossRefGoogle Scholar
ZAHLER, M., ESSIG, A., GOTHE, R. & RINDER, H. (1999). Molecular analyses suggest monospecificity of the genus Sarcoptes (Acari: Sarcoptidae). International Journal for Parasitology 29, 759766.CrossRefGoogle Scholar