Molecular cloning and allergenicity study of tropomyosin from shrimp commonly consumed in Thailand

Name: Panyarat Laurchan

LCSH: Kasetsart University -- Theses. M.S. (Biochemistry) 2020

Classification :.LCCS: QP552.M93

LCSH: Kasetsart University. -- Department of Biochemistry -- Theses

LCSH: Tropomyosins

LCSH: Food allergy

LCSH: Allergy

LCSH: Shrimps

LCSH: Immunoglobulin E

Abstract: Allergy is a type I hypersensitivity reaction, which is mediated by immunoglobulin E (IgE) antibodies and stimulated by a number of substances including food. Upon consuming food containing allergens, IgE molecules are produced and stimulate allergic reactions varied from rash, red eyes, runny nose, diarrhoea, trouble breathing or even death. Crustacean is group of the most common allergenic food in both children and adults. The major protein allergen mediated allergic reaction of crustaceans is tropomyosin. Tropomyosin is a highly heat-stable protein, which is an alpha-helical coiled-coil homo-dimeric structure. The amino acid sequences of tropomyosin are highly conserved among the crustaceans. Therefore, the high sequence identity of tropomyosin induces IgE cross-reactivity among crustacean species. However, this cross-reactivity toward different crustacean species in Thailand is not well studied. To gain more information, this study aimed to identify the cross reactivity and clone tropomyosin from 8 species of crustacean that are commonly consumed in Thailand. The crude proteins were extracted from crustacean with extraction buffer at different pH and the total protein was measured. The protein profiles were compared by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). Protein allergens were probed using pooled volunteers’ sera with a history of crustacean allergy and analysed by immunoblotting, LC-MS/MS and immunoprecipitation (IP). To identify the amino acid sequence of tropomyosin, the cDNA of tropomyosin was cloned from the muscle of the other crustacean by reverse-transcription polymerase chain reaction. The result showed that the protein profiles of each species were dependent on the pH of the extraction buffer. The immunoblotting, LC-MS/MS and IP analysis revealed that the cross-reactivity among 8 species of crustacean was responsible by a few groups of proteins including tropomyosin. The amino acid sequences of tropomyosins were highly conserved among crustaceans, which suggested that tropomyosin play important role in cross-reactivity among crustacean. In conclusion, this study provided more information toward understating of crustacean allergy. These data can be further used to predict cross-species allergy, to develop allergy testing and to develop a test kit for detecting allergen contamination in the food industry.

Kasetsart University. Office of the University Library

Address: Bangkok

Email: tdckulib@ku.ac.th

Name: Chomdao Sinthuvanich

Role: Thesis Advisor

Name: Pharima Phiriyangkul

Role: Thesis CO-Advisor

Created: 2020

Modified: 2025-07-18

Issued: 2025-07-18

วิทยานิพนธ์/Thesis

application/pdf

URL: https://www.lib.ku.ac.th/KUthesis/2563/panyarat-lau-all.pdf

CallNumber: QP552.M93 .P36

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biochemistry

Grantor: Kasetsart University

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