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Isolation, cDNA cloning and expression of Lig v 1, the major allergen from privet pollen.
Clin Exp Allergy. 1996 Dec; 26(12):1401-10.CE

Abstract

BACKGROUND

An olive allergen-like protein has been detected in privet pollen. This protein could be involved in the allergenic cross-reactivity described for privet and olive tree pollen extracts.

OBJECTIVE

Isolation and characterization of natural Lig v 1. Cloning and expression of its cDNA in order to assess its structural similarity with the olive allergen.

METHODS

Current chromatographic methods were used to isolate the privet counterpart of Ole e 1. A pool of sera from subjects allergic to olive tree pollen was used to immunodetect the protein in the elution profiles. Ole e 1-specific polyclonal antibody and allergic sera were used in immunoblotting assays of the isolated protein. Polymerase chain reaction amplification of the first strand cDNA synthesized from the privet pollen total RNA was carried out to prepare a full-length fragment encoding Lig v 1. After nucleotide sequencing, expression of one clone was performed in Escherichia coli, under the form of a fusion protein with glutathione S-transferase. The IgE binding capability of the recombinant protein was also analysed.

RESULTS

The major allergen from privet pollen. Lig v 1, was purified to homogeneity by two gel filtration chromatographies and one reverse-phase high-performance liquid chromatography. Its amino acid composition and N-terminal amino acid sequence were determined. Two different clones encoding Lig v 1 were sequenced. Strong sequence similarity between Lig v 1 and Ole e 1 was observed, the identity being 85 and 96%. One of the sequenced clones was expressed and the recombinant product exhibited IgG and IgE binding activities against both anti-Ole e 1 polyclonal antibodies and olive-allergic sera.

CONCLUSION

Privet pollen contains a protein structurally and immunologically related to the major allergen of olive pollen. The similarity exhibited by these proteins could explain the cross-reactivity observed between the two pollen extracts. Since these allergens are highly polymorphic, the expression of an immunologically active recombinant Lig v 1 will permit the preparation of well defined molecules for both research and clinical purposes.

Authors+Show Affiliations

Departamento de Bioquímica y Biología Molecular, Facultad de Química, Universidad Complutense, Madrid, Spain.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

9027441

Citation

Batanero, E, et al. "Isolation, cDNA Cloning and Expression of Lig V 1, the Major Allergen From Privet Pollen." Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology, vol. 26, no. 12, 1996, pp. 1401-10.
Batanero E, Gonzalez De La Peña MA, Villalba M, et al. Isolation, cDNA cloning and expression of Lig v 1, the major allergen from privet pollen. Clin Exp Allergy. 1996;26(12):1401-10.
Batanero, E., Gonzalez De La Peña, M. A., Villalba, M., Monsalve, R. I., Martin-Esteban, M., & Rodríguez, R. (1996). Isolation, cDNA cloning and expression of Lig v 1, the major allergen from privet pollen. Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology, 26(12), 1401-10.
Batanero E, et al. Isolation, cDNA Cloning and Expression of Lig V 1, the Major Allergen From Privet Pollen. Clin Exp Allergy. 1996;26(12):1401-10. PubMed PMID: 9027441.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation, cDNA cloning and expression of Lig v 1, the major allergen from privet pollen. AU - Batanero,E, AU - Gonzalez De La Peña,M A, AU - Villalba,M, AU - Monsalve,R I, AU - Martin-Esteban,M, AU - Rodríguez,R, PY - 1996/12/1/pubmed PY - 1996/12/1/medline PY - 1996/12/1/entrez SP - 1401 EP - 10 JF - Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology JO - Clin Exp Allergy VL - 26 IS - 12 N2 - BACKGROUND: An olive allergen-like protein has been detected in privet pollen. This protein could be involved in the allergenic cross-reactivity described for privet and olive tree pollen extracts. OBJECTIVE: Isolation and characterization of natural Lig v 1. Cloning and expression of its cDNA in order to assess its structural similarity with the olive allergen. METHODS: Current chromatographic methods were used to isolate the privet counterpart of Ole e 1. A pool of sera from subjects allergic to olive tree pollen was used to immunodetect the protein in the elution profiles. Ole e 1-specific polyclonal antibody and allergic sera were used in immunoblotting assays of the isolated protein. Polymerase chain reaction amplification of the first strand cDNA synthesized from the privet pollen total RNA was carried out to prepare a full-length fragment encoding Lig v 1. After nucleotide sequencing, expression of one clone was performed in Escherichia coli, under the form of a fusion protein with glutathione S-transferase. The IgE binding capability of the recombinant protein was also analysed. RESULTS: The major allergen from privet pollen. Lig v 1, was purified to homogeneity by two gel filtration chromatographies and one reverse-phase high-performance liquid chromatography. Its amino acid composition and N-terminal amino acid sequence were determined. Two different clones encoding Lig v 1 were sequenced. Strong sequence similarity between Lig v 1 and Ole e 1 was observed, the identity being 85 and 96%. One of the sequenced clones was expressed and the recombinant product exhibited IgG and IgE binding activities against both anti-Ole e 1 polyclonal antibodies and olive-allergic sera. CONCLUSION: Privet pollen contains a protein structurally and immunologically related to the major allergen of olive pollen. The similarity exhibited by these proteins could explain the cross-reactivity observed between the two pollen extracts. Since these allergens are highly polymorphic, the expression of an immunologically active recombinant Lig v 1 will permit the preparation of well defined molecules for both research and clinical purposes. SN - 0954-7894 UR - https://www.unboundmedicine.com/medline/citation/9027441/Isolation_cDNA_cloning_and_expression_of_Lig_v_1_the_major_allergen_from_privet_pollen_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0954-7894&date=1996&volume=26&issue=12&spage=1401 DB - PRIME DP - Unbound Medicine ER -