Premium
Molecular cloning and functional expression of allergenic sarcoplasmic calcium‐binding proteins from Penaeus shrimps
Author(s) -
Mita Hajime,
Koketsu Aiko,
Ishizaki Shoichiro,
Shiomi Kazuo
Publication year - 2013
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.5961
Subject(s) - shrimp , penaeus monodon , biology , crustacean , complementary dna , cloning (programming) , penaeus , molecular cloning , biochemistry , fishery , gene , computer science , programming language
BACKGROUND Sarcoplasmic calcium‐binding proteins ( SCPs ) have recently been identified as crustacean allergens. However, information on their primary structures is very limited and no recombinant SCP ( rSCP ) as an alternative of natural SCP ( nSCP ) is available. This study was aimed to elucidate primary structures of SCPs from two species of Penaeus shrimp (black tiger shrimp and kuruma shrimp) by cDNA cloning and to produce a black tiger shrimp rSCP preparation that is comparable in IgE reactivity to nSCP .RESULTS The full‐length cDNAs encoding black tiger shrimp and kuruma shrimp SCPs were successfully cloned. Both SCPs are composed of 193 amino acid residues and share more than 80% sequence identity with the known crustacean SCPs . The black tiger shrimp SCP was then expressed in Escherichia coli using the pFN6A ( HQ ) Flexi vector system. Enzyme‐linked immunosorbent assay ( ELISA ) and inhibition ELISA experiments demonstrated that rSCP has the same IgE reactivity as nSCP .CONCLUSION Our results provide further evidence for the high sequence identity among crustacean SCPs . In addition, rSCP will be a useful tool in studying crustacean allergens and also in the diagnosis of crustacean allergy. © 2012 Society of Chemical Industry