G. mellonella_TimeDependent_Proteome
Description
Thermal variation significantly affects insect physiology, cellular balance, and stress resilience. In this study, we conducted a comparative proteomic analysis of Galleria mellonella larvae exposed to three temperatures (18, 30, and 37 °C) for 24 and 48 hours to understand the molecular responses over time to thermal stress. Twodimensional gel electrophoresis followed by LC–MS/MS identified distinct sets of proteins uniquely expressed at each temperature–time combination, as well as a core group consistently present across treatments.
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Steps to reproduce
Galleria mellonella larvae were reared under controlled laboratory conditions and subsequently subjected to temperature treatments at the specified experimental temperatures (18, 30, and 37 °C) and periods (24 and 48 h). Following exposure, proteins were isolated from hemolymph using a trichloroacetic acid (TCA)-based precipitation protocol. Protein concentrations were quantified with the Pierce 660 nm Protein Assay Kit according to the manufacturer’s instructions. Proteomic profiles were generated using two-dimensional gel electrophoresis (2D-PAGE), and gel images were comparatively analyzed. Selected spots were manually excised from the gels and processed for mass spectrometric identification. Briefly, excised gel pieces were prepared using standard in-gel digestion procedures before analysis by liquid chromatography–tandem mass spectrometry (LC–MS/MS). The subcellular localization of the identified proteins was predicted using WoLF PSORT, and their functional classification was performed based on Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotations.
Institutions
- Erciyes Universitesi