Purification, cloning, expression and immunological analysis of XXXX @entity369 kinase, the crab allergen.

@entity8389 @entity535 @entity566 @entity56 @entity54558 @entity13724 @entity369
BACKGROUND: Although crustaceans have been reported to be one of the most common causes of IgE-mediated @entity56 , there are no reports about the characterization and identification of @entity369 kinase ( @entity54558 ) from the @entity8389 ( @entity8389 ) as allergen. In the present study, the purification, molecular cloning, expression and immunological analyses of the IgE allergen @entity54558 from the @entity8389 were investigated. RESULTS: The results showed that cloned DNA fragments of @entity54558 from the @entity8389 had open reading frames of 1021 bp, predicted to encode proteins with 356 @entity535 residues. Sequence alignment revealed that @entity8389 @entity54558 shares high homology with other crustacean species. @entity8389 @entity54558 gene was further recombined with the vector of pGEX-4T-3 and expressed in @entity13724 . 2-D electrophoresis suggested that native @entity54558 (nAK) and recombinant @entity54558 ( @entity54558 ) shared the same molecular weight of 40 kDa, and the pI is 6.5 and 6.3, respectively. The nAK and @entity54558 were further confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Immunoblotting analysis and colloidal gold immunochromatographic assay (GICA) using sera from subjects with crustacean @entity566 confirmed that the nAK and @entity54558 reacted positively with these sera, indicating @entity54558 is a specific allergen of @entity8389 . CONCLUSION: Both of purified nAK and @entity54558 reacted positively with sera from subjects with crustacean @entity566 in immunoblotting and GICA analysis, indicating @entity54558 is a common allergen of @entity8389 . In vitro expressed @entity54558 is proposed as a source of the protein for immunological or clinical studies.
