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Type :article
Subject :Q Science (General)
ISSN :0301-4851
Main Author :Rosmilah Misnan
Additional Authors :Nur Farah Hani Azemi
Poh, Keong Bun
Marina Mokhta
Nurhaida Kamaruddin
Fah, Wongchee
Title :Molecular and allergenic characterization of recombinant tropomyosin from mud crab Scylla olivacea
Place of Production :Tanjung Malim
Publisher :Fakulti Sains dan Matematik
Year of Publication :2021
Notes :Molecular Biology Reports
Corporate Name :Universiti Pendidikan Sultan Idris
HTTP Link :Click to view web link

Abstract : Universiti Pendidikan Sultan Idris
Background: Tropomyosin is a major allergen in crustaceans, including mud crab species, but its molecular and allergenic properties in Scylla olivacea are not well known. Thus, this study aimed to produce the recombinant tropomyosin protein from S. olivacea and subsequently investigate its IgE reactivity. Methods and Results: The tropomyosin gene was cloned and expressed in the Escherichia coli system, followed by SDS-PAGE and immunoblotting test to identify the allergenic potential of the recombinant protein. The 855-base pair of tropomyosin gene produced was found to be 99.18% homologous to Scylla serrata. Its 284 amino acids matched the tropomyosin of crustaceans, arachnids, insects, and Klebsiella pneumoniae, ranging from 79.03 to 95.77%. The tropomyosin contained 89.44% alpha-helix folding with a tertiary structure of two-chain alpha-helical coiled-coil structures comprising a homodimer heptad chain. IPTG-induced histidine tagged-recombinant tropomyosin was purified at the size of 42�kDa and confirmed as tropomyosin using anti-tropomyosin monoclonal antibodies. The IgE binding of recombinant tropomyosin protein was reactive in 90.9% (20/22) of the sera from crab-allergic patients. Conclusions: This study has successfully produced an allergenic recombinant tropomyosin from S. olivacea. This recombinant tropomyosin may be used as a specific allergen for the diagnosis of allergy. ? 2021, The Author(s), under exclusive licence to Springer Nature B.V.

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