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Yeast two hybrid y2h assay

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The Yeast two-hybrid (Y2H) assay is a laboratory technique used to detect and study protein-protein interactions. It allows for the identification of direct physical interactions between two proteins of interest expressed in yeast cells.

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Lab products found in correlation

Easy yeast plasmid isolation kit, by Takara Bio (1 mentions) Smart cdna library construction kit, by Takara Bio (1 mentions) Yeast maker transformation system 2 kit, by Takara Bio (1 mentions) Rna extraction kit, by Aidlab (1 mentions)

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Y2H assay (19 citations) Yeast two-hybrid assay (12 citations)

2 protocols using yeast two hybrid y2h assay

1

Yeast Two-Hybrid Assay Protocol

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The OseIF6.1 and OsNMD3 CDS have been cloned into plasmids pGBKT7 and pGADT7, respectively. Ligation-independent cloning (LIC) was used to construct all plasmids (Aslanidis et al., 1994 (link)). The Yeast two-hybrid (Y2H) assay was conducted in accordance with the manufacturer’s instructions (Clontech). The pGBKT7-53 plasmid was transformed with pGADT7-T as a positive control, and the pGBKT7-Lam plasmid was transformed with pGADT7-T into the Y2HGold strain as a negative control. All the primers used are listed in Supplementary Table 1.
Guo H., Lv J., Su X., Chen L., Ren J., Liu L., Ren M., Liu S., Dai M., Ren G, & Gao F. (2024). Rice OseIF6.1 encodes a eukaryotic translation initiation factor and is essential for the development of grain and anther. Frontiers in Plant Science, 15, 1366986.
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2

Construction and Screening of cDNA Library

Cited 1 time
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The cDNA library was constructed as previously described by Ding et al. [20 (link)]. The total mRNA was isolated by an RNA Extraction Kit (Aidlab Biotechnologies Co., Ltd., Beijing, China). The cDNA library was constructed using mixed RNA samples of fruit stored at 7 °C (1, 2, 3, 6, 9, 12 days) by the SMART™ cDNA Library Construction Kit (Clontech, Cat. No. 634901, Fitchburg, WI, USA). The yeast transformation was done using a Yeast Maker Transformation System 2 Kit (Clontech, Cat. NO. 630439). Yeast two-hybrid (Y2H) assay (Clontech, Cat. No. 630489) was conducted according to the manufacturer’s instructions. Yeast plasmid was extracted using an Easy Yeast Plasmid Isolation Kit (Clontech, Cat. No. 630467).
Song Z., Qin J., Zheng Q., Ding X., Chen W., Lu W., Li X, & Zhu X. (2019). The Involvement of the Banana F-Box Protein MaEBF1 in Regulating Chilling-Inhibited Starch Degradation through Interaction with a MaNAC67-Like Protein. Biomolecules, 9(10), 552.
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