In Vivo Monitoring of Transcriptional Activity During Metabolic Transition Using a Bioluminescent Reporter in Yeast.

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Tác giả: José Benavides-Parra, Francisco A Cubillos, Luis F Larrondo, Felipe Muñoz-Guzmán, Catalina Muñoz-Tapia, Pablo Quintrel

Ngôn ngữ: eng

Ký hiệu phân loại: 949.59012 *Greece

Thông tin xuất bản: United States : Journal of visualized experiments : JoVE , 2025

Mô tả vật lý:

Bộ sưu tập: NCBI

ID: 684660

Sequential sugar consumption, from a preferred sugar source to a less preferred one, represents a critical metabolic adaptation in yeast, which is particularly relevant for survival in fluctuating environments such as those found in beer fermentation. However, sugar transitions are an environmental variable that is challenging to predict and detect, impacting the outcome of beer fermentations. This protocol describes an in vivo system to monitor transcriptional activation associated with the glucose-to-maltose metabolic shift in Saccharomyces eubayanus that applies to different wild Saccharomyces yeast strains. The system employs an episomal bioluminescent transcriptional reporter for maltose metabolism, focusing on MAL32, since it provides a good readout for metabolic shifts, as studied in S. cerevisiae. For this, yeast strains were transformed with plasmids containing the MAL32 regulatory region from S. eubayanus, controlling the expression of a gene encoding for a destabilized version of firefly luciferase
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