This study aims to characterize the molecular and allergenic properties of tropomyosin from local mud crab Scylla species. The methodology involved quantifying reference genes and tropomyosin levels in these crabs using real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Additionally, recombinant tropomyosin gene and protein were created using recombinant DNA techniques. The reactivity of tropomyosin with IgE antibodies was assessed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDSPAGE) and anti-tropomyosin monoclonal antibody immunoblotting. Bioinformatic tools were used to predict various characteristics such as physicochemical properties, structural composition, phylogenetic relationships, IgE-epitope regions, and docking simulations of the recombinant proteins. The findings indicated that elongation factor 1 alpha (EF1_) was a suitable normalizer for qRT-PCR in the three mud crab species studied, and a combination of three reference.....

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