Yeast One-Hybrid Assay for Transcriptional Interactions

Y1H assays were carried out using the Matchmaker Gold Yeast One-Hybrid System (Clontech, Fitchburg, WI, USA). The promoters of MaSGR1, MaGWD1, MaAMY3, MaPL8, MaEXP-A8, and MaXYL23-like were cloned into pAbAi, which was stored in our laboratory. It was then linearized and separately introduced into the Y1H Gold strain. The CDS of MaERF012 was ligated to the pGADT7 vector and subsequently introduced into the yeast strain. It was then transferred into the aforementioned bait reporter yeast strain containing the promoters of MaSGR1, MaGWD1, MaAMY3, MaPL8, MAEXP-A8, and MaXYL23-like. Their interaction was tested according to the method of Song et al. [12 (link)].

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Chen H., Lai X., Wang L., Li X., Chen W., Zhu X, & Song Z. (2022). Ethylene Response Factor MaERF012 Modulates Fruit Ripening by Regulating Chlorophyll Degradation and Softening in Banana. Foods, 11(23), 3882.

Publication 2022

Matchmaker Gold Yeast One-Hybrid System

Assays Gold Hybrid Strain Vector Yeast

Corresponding Organization : Shandong Agricultural University

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Variable analysis

independent variables

Promoters of MaSGR1, MaGWD1, MaAMY3, MaPL8, MaEXP-A8, and MaXYL23-like genes
CDS of MaERF012 gene

dependent variables

Interaction between MaERF012 and the promoters of MaSGR1, MaGWD1, MaAMY3, MaPL8, MaEXP-A8, and MaXYL23-like genes

control variables

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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