Gene Cloneing, Expression and Immunological Identification of Main Allergen Gal d 3 in Eggs

doi:10.7506/spkx1002-6630-200901034

FOOD SCIENCE ›› 2009, Vol. 30 ›› Issue (1): 142-146.doi: 10.7506/spkx1002-6630-200901034

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Gene Cloneing, Expression and Immunological Identification of Main Allergen Gal d 3 in Eggs

WU Xu-li，YANG Zhi-hua，LIU Zhi-gang*，WU Hai-qiang

(Institute of Allergy and Immunology, Shenzhen University, Shenzhen 518060, China)

Received:2007-12-06 Revised:2008-04-14 Online:2009-01-01 Published:2010-12-29
Contact: LIU Zhi-gang E-mail:lzg@szu.edu.cn

Abstract

Abstract:

Objective: To clone, express and identify the gene of Gal d 3 in eggs. Methods: The total RNA was extracted from eggs, and the gene of Gal d 3 was amplified by RT-PCR and cloned into pMD18-T for sequencing. Then the interesting gene was subcloned into pET-28a for expression. After purification of the recombinant protein, its IgE-binding capacity was determined by Western blotting. Results: The gene of Gal d 3 in eggs is obtained by clone, which has 2118 bp and encodes 705 amino acids. This isoelectric point of the encoded protein is 6.5 with a molecular weight of 76000. Conclusion: The main allergen gene Gal d 3 in eggs is successfully cloned and expressed in this study, and this recombinant protein shows good IgE-binding capacity.

Key words: eggs, allergen, clone, expression, Gal d 3

CLC Number:

WU Xu-li，YANG Zhi-hua，LIU Zhi-gang*，WU Hai-qiang. Gene Cloneing, Expression and Immunological Identification of Main Allergen Gal d 3 in Eggs[J]. FOOD SCIENCE, 2009, 30(1): 142-146.

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Gene Cloneing, Expression and Immunological Identification of Main Allergen Gal d 3 in Eggs

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