What is the protocol of the yeast two hybrid system?
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The yeast two-hybrid system protocol involves several key steps. Firstly, library yeast is activated, and an AD-Y fusion vector is transformed into the library yeast . Secondly, bait plasmids BD-X are transformed into yeast cells Y2H Gold . Thirdly, recombined yeast of BD-X and AD-Y are fused into a diploid, cultured on an amplification medium with zinc ions, and positive clone screening is conducted . This method utilizes GAL4 protein, a zinc ion-dependent transcription factor, to enhance interactions between low-abundance proteins, improving the detection rate of low-interaction proteins . The protocol is versatile, suitable for simple transformations, targeting in yeast, GAP repair, and multi-plasmid transformations for two-hybrid systems .
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| Papers (5) | Insight |
|---|---|
| The yeast two-hybrid system is utilized for mapping novel protein interactions, aiding in identifying viral-host interactions crucial for disease pathogenesis and potential antiviral therapies. | |
| The yeast two-hybrid method involves activating library yeast, transforming AD-Y fusion vector, introducing bait plasmids BD-X, fusing recombined yeast, culturing in zinc-containing medium, and conducting positive clone screening. | |
| The protocol involves screening ORFs using minipools, verifying positives, sequencing for interaction pairs, and final verification with fresh inoculants, ensuring low false-discovery rates for high-quality network mapping. | |
| The yeast transformation protocol v1 is suitable for multi-plasmid transformations for two-hybrid systems, as outlined in Gietz and Woods (2002) and may require modifications for specific needs. | |
| The yeast transformation protocol based on Gietz and Woods (2002) includes steps for simple transformation, targeting, GAP repair, and multi-plasmid transformations for the yeast two-hybrid system. |
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