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Cloning and sequencing of a cDNA expressing a ribosomal P0 peptide from Culicoides nubeculosus (Diptera).
Vet Immunol Immunopathol. 2004 May; 99(1-2):99-111.VI

Abstract

Insect bite dermal hypersensitivity (IBH) is an allergic dermatitis of horses caused by bites of Culicoides spp. and sometimes Simulium spp. The aim of the investigation presented here was to identify allergens causing IBH. A cDNA library expressing recombinant Culicoides nubeculosus proteins was screened using affinity-purified serum from an IBH-affected horse. Screening of the library resulted in identification of one immunoreactive clone. The sequence of the cDNA insert was determined and revealed a 600 bp insert with an open reading frame coding for a 78 amino acid long protein, called rCul n 1. Analysis of the deduced amino acid sequence revealed an identity of 67-78% to the C-terminal part of the 318 amino acid long ribosomal P0 protein from other Diptera. Furthermore, the 38 C-terminal amino acids displayed an identity of 57% with the C-terminal part of the acidic ribosomal protein P2 from Aspergillus fumigatus. The cDNA insert was subcloned and expressed as a [His]6-tagged protein in Escherichia coli and purified using Ni2(+)-chelate affinity chromatography. The 10kDa recombinant Cul n 1 protein bound the affinity-purified antibody fraction used for screening the expression library. Determination of IgE and IgG levels against rCul n 1 by ELISA in sera from 19 IBH-affected and 18 Swiss control horses and in sera from eight control horses living in Iceland showed no significant differences between the three groups of horses (median IgE levels = 60, 49 and 44 relative ELISA units, respectively). rCul n 1 did not induce sulfidoleukotriene (sLT) release from peripheral blood leukocytes of IBH-affected horses (N = 5), although sLT release was induced with the Culicoides whole body extract.

Authors+Show Affiliations

Division of Immunogenetics, Institute of Animal Genetics, Nutrition and Housing, Bremgartenstrasse 109 A, 3012-Berne, Switzerland.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15113658

Citation

Althaus, H, et al. "Cloning and Sequencing of a cDNA Expressing a Ribosomal P0 Peptide From Culicoides Nubeculosus (Diptera)." Veterinary Immunology and Immunopathology, vol. 99, no. 1-2, 2004, pp. 99-111.
Althaus H, Müller N, Busato A, et al. Cloning and sequencing of a cDNA expressing a ribosomal P0 peptide from Culicoides nubeculosus (Diptera). Vet Immunol Immunopathol. 2004;99(1-2):99-111.
Althaus, H., Müller, N., Busato, A., Mellor, P. S., Torsteinsdottir, S., & Marti, E. (2004). Cloning and sequencing of a cDNA expressing a ribosomal P0 peptide from Culicoides nubeculosus (Diptera). Veterinary Immunology and Immunopathology, 99(1-2), 99-111.
Althaus H, et al. Cloning and Sequencing of a cDNA Expressing a Ribosomal P0 Peptide From Culicoides Nubeculosus (Diptera). Vet Immunol Immunopathol. 2004;99(1-2):99-111. PubMed PMID: 15113658.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cloning and sequencing of a cDNA expressing a ribosomal P0 peptide from Culicoides nubeculosus (Diptera). AU - Althaus,H, AU - Müller,N, AU - Busato,A, AU - Mellor,P S, AU - Torsteinsdottir,S, AU - Marti,E, PY - 2002/10/24/received PY - 2003/12/12/revised PY - 2004/01/19/accepted PY - 2004/4/29/pubmed PY - 2004/7/28/medline PY - 2004/4/29/entrez SP - 99 EP - 111 JF - Veterinary immunology and immunopathology JO - Vet Immunol Immunopathol VL - 99 IS - 1-2 N2 - Insect bite dermal hypersensitivity (IBH) is an allergic dermatitis of horses caused by bites of Culicoides spp. and sometimes Simulium spp. The aim of the investigation presented here was to identify allergens causing IBH. A cDNA library expressing recombinant Culicoides nubeculosus proteins was screened using affinity-purified serum from an IBH-affected horse. Screening of the library resulted in identification of one immunoreactive clone. The sequence of the cDNA insert was determined and revealed a 600 bp insert with an open reading frame coding for a 78 amino acid long protein, called rCul n 1. Analysis of the deduced amino acid sequence revealed an identity of 67-78% to the C-terminal part of the 318 amino acid long ribosomal P0 protein from other Diptera. Furthermore, the 38 C-terminal amino acids displayed an identity of 57% with the C-terminal part of the acidic ribosomal protein P2 from Aspergillus fumigatus. The cDNA insert was subcloned and expressed as a [His]6-tagged protein in Escherichia coli and purified using Ni2(+)-chelate affinity chromatography. The 10kDa recombinant Cul n 1 protein bound the affinity-purified antibody fraction used for screening the expression library. Determination of IgE and IgG levels against rCul n 1 by ELISA in sera from 19 IBH-affected and 18 Swiss control horses and in sera from eight control horses living in Iceland showed no significant differences between the three groups of horses (median IgE levels = 60, 49 and 44 relative ELISA units, respectively). rCul n 1 did not induce sulfidoleukotriene (sLT) release from peripheral blood leukocytes of IBH-affected horses (N = 5), although sLT release was induced with the Culicoides whole body extract. SN - 0165-2427 UR - https://www.unboundmedicine.com/medline/citation/15113658/Cloning_and_sequencing_of_a_cDNA_expressing_a_ribosomal_P0_peptide_from_Culicoides_nubeculosus__Diptera__ L2 - https://linkinghub.elsevier.com/retrieve/pii/S016524270400025X DB - PRIME DP - Unbound Medicine ER -