Identification and characterisation of the major allergen of Chinese mitten crab (Eriocheir sinensis)

Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn)

“…Then 50 毺g of prawn extract was applied to immobilized non-linear pH [3][4][5][6][7][8][9][10] gradient strip of 7 cm length for rehydration overnight (12)(13)(14) h). Isoelectric focusing was run at 20 曟 using the Protean IEF Cell Apparatus (BioRad, USA) with the following voltage/time gradient: 100 V for 1 min, 250 V for 30 min, 4 000 V for 2 h and 4 000 V for 10 000 V-h. Before transferring the IPG strip onto the second dimension, the strip was equilibrated sequentially for 10 min in a buffer containing 65 mM dithio-threitol and then 135 mM iodoacetamide in 125 mM Tris-HCl, pH 6.8, 6 M urea, 2% SDS, 30% glycerol and 0.01% bromophenol blue.…”

“…In addition, tropomyosin has also been identified as the major allergen of other crustaceans [9,10] , mollusks [11][12][13][14][15] , house dust mites [16,17] and cockroaches [18,19] .…”

Purification, cloning, expression and immunological analysis of Scylla serrata arginine kinase, the crab allergen

“…Isolation of RNA, cDNA cloning, and sequencing Isolation of RNA, cDNA cloning, and sequencing procedures were performed according to Liang et al 15 Total RNA was extracted from fresh muscle with Trizol reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer's protocol. Single-stranded CDNA was synthesized with reverse transcriptase (Invitrogen) and an oligo(dT) 15 primer, according to the manufacturer's instructions and this CDNA was used as a template for reverse transcription polymerase chain reaction (RT-PCR).…”

“…14 Among shellfish, crab is assumed to be mainly responsible for severe hypersensitive reactions in China, as there are more than 600 kinds of crab in China, including Chinese mitten crab (Eriocheir sinensis), mud crab and swimming crab, etc. 15 Mud crab (Scylla serrata), which is widely distributed in China, is an economically important fished and farmed crab species in many areas of Asia.…”

Characterization of Seafood Proteins Causing Allergic Diseases

“…The most common seafood allergens and the used method(s) for the characterization and quantification are summarized in Table 2. , 2010b;Huang et al, 2010;Kunimoto et al, 2009;Leung & Chu, 1998;Liang et al, 2008;Motoyama et al, 2007;Shibahara et al, 2009;Werner et al, 2007;Yu et al, 2010) Crab DeWitt et al, 2004;Leung et al, 1996;Leung & Chu, 1998;Motoyama et al, 2007;Mykles et al, 1998;Shibahara et al, 2009;Werr et al, 2009) To characterize seafood allergens, it is necessary to profile the complex crude extracts by separating the protein content by gel electrophoresis. This technique plays a central role in seafood allergen research, wherein it provides a powerful separation and semi-quantification information.…”