CLONING, EXPRESSION, AND PRIMARY STRUCTURE OF METAPENAEUS-ENSIS TROPOMYOSIN, THE MAJOR HEAT-STABLE SHRIMP ALLERGEN

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY

卷:94

Article

LEUNG, PSC ; CHU, KH ; CHOW, WK ; ANSARI, A ; BANDEA, CI ; KWAN, HS ; NAGY, SM ; GERSHWIN, ME

关键词: SHRIMP ALLERGY;    METAPENAEUS ENSIS;    IGE REACTIVITY;    CDNA CLONE;    TROPOMYOSIN;    RECOMBINANT PROTEIN;    EPITOPE;   

DOI  :  10.1016/0091-6749(94)90156-2

来源: Elsevier

【 摘 要 】

Shrimp is a common cause of seafood hypersensitivity. To study the mechanism of seafood hypersensitivity at the molecular level, we have determined the primary structure of the major heat-stable allergen of shrimp by cloning, expression, nucleotide sequencing, and amino acid sequence determination of an IgE-reactive cDNA clone, Met e I, isolated from a Metapenaeus ensis expression library in lambda gt 11. We first constructed a cDNA library from the shrimp M. ensis in lambda gt 11. We then screened the library with sera from patients with hypersensitivity reactions to shrimp and identified a positive IgE-reactive clone, designated as Met e I. This cDNA was purified to homogeneity and subsequently expressed in the plasmid pGEX. Serum antibodies from patients with shrimp allergy demonstrated positive IgE reactivity by immunoblotting to a protein encoded by the clone Met e I; sera from nonallergic control subjects were not reactive. The nucleotide sequence of this cDNA clone revealed art open reading frame of 281 amino acid residues, coding for a protein of 34 kd. Comparison of the Met e I amino acid sequence with the Genbank database showed that Met e I is highly homologous to multiple isoforms of tropomyosin.

【 授权许可】

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