title: Molecular cloning and functional expression of allergenic sarcoplasmic calcium-binding proteins from Penaeus shrimps.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Minato-ku, Tokyo, 108-8477, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Mita
authors.forename: Aiko
authors.initials: A
authors.lastname: Koketsu
authors.forename: Shoichiro
authors.initials: S
authors.lastname: Ishizaki
authors.forename: Kazuo
authors.initials: K
authors.lastname: Shiomi
mesh_terms: D000485:Allergens; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary; D004032:Diet; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D005512:Food Hypersensitivity / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism; D033561:Penaeidae / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017385:Sequence Homology; D012758:Shellfish; D013045:Species Specificity
chemical_list: D000485:Allergens; D000596:Amino Acids; D002135:Calcium-Binding Proteins; D018076:DNA, Complementary; D011994:Recombinant Proteins; D007073:Immunoglobulin E