Podcast
Questions and Answers
Protein-protein interactions can be classified based on ______
Protein-protein interactions can be classified based on ______
composition
______ proteins carry out their functions as homo-oligomers
______ proteins carry out their functions as homo-oligomers
Several enzymes, carrier proteins, scaffolding proteins, and transcriptional regulatory factors
Targeted modulation of PPIs with small molecules is a promising approach in ______ development
Targeted modulation of PPIs with small molecules is a promising approach in ______ development
drug
The yeast 2-hybrid assay is a technique used to detect ______ interactions
The yeast 2-hybrid assay is a technique used to detect ______ interactions
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Yeast two-hybrid experiment detects the physical interactions of proteins through the activation of a ______ gene
Yeast two-hybrid experiment detects the physical interactions of proteins through the activation of a ______ gene
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Most commonly, transcription in a yeast two-hybrid experiment is measured by ______ selection
Most commonly, transcription in a yeast two-hybrid experiment is measured by ______ selection
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The Y2H assay relies on the expression of a reporter gene (such as lacZ or GFP), which is activated by the binding of a particular ______ factor.
The Y2H assay relies on the expression of a reporter gene (such as lacZ or GFP), which is activated by the binding of a particular ______ factor.
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The query protein of interest fused with the BD is known as the ______, and the protein library fused with the AD is referred to as the Prey.
The query protein of interest fused with the BD is known as the ______, and the protein library fused with the AD is referred to as the Prey.
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In order to activate the reporter gene expression, a ______ unit must be present at the gene locus.
In order to activate the reporter gene expression, a ______ unit must be present at the gene locus.
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Expression of a reporter gene requires the binding of a transcription factor, which normally consists of two functionally and structurally independent domains: DNA-binding (DB) and ______ domains.
Expression of a reporter gene requires the binding of a transcription factor, which normally consists of two functionally and structurally independent domains: DNA-binding (DB) and ______ domains.
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In the absence of Bait-Prey interaction, the AD domain is unable to localize to the reporter gene to drive gene ______.
In the absence of Bait-Prey interaction, the AD domain is unable to localize to the reporter gene to drive gene ______.
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However, when Bait and Prey interact, the DB domain binds to the DNA localizing the AD domain upstream of the reporter gene, leading to the expression of reporter ______.
However, when Bait and Prey interact, the DB domain binds to the DNA localizing the AD domain upstream of the reporter gene, leading to the expression of reporter ______.
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Advantages of Y2H: Y2H is a relatively fast and easy way to screen for ______ interactions
Advantages of Y2H: Y2H is a relatively fast and easy way to screen for ______ interactions
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Y2H requires little hands-on time and technical skill, making it ______ to use
Y2H requires little hands-on time and technical skill, making it ______ to use
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Y2H can be scaled up by screening yeast libraries of tagged “prey” proteins against a single “bait”, allowing thousands of potential interactions to be screened ______
Y2H can be scaled up by screening yeast libraries of tagged “prey” proteins against a single “bait”, allowing thousands of potential interactions to be screened ______
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Limitations of Yeast Two-Hybrids: There are high rates of false positives and false negatives in Y2H ______
Limitations of Yeast Two-Hybrids: There are high rates of false positives and false negatives in Y2H ______
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Protein folding, expression, and post-translational modifications may not be the same in yeast as they are in humans, affecting their physical ______
Protein folding, expression, and post-translational modifications may not be the same in yeast as they are in humans, affecting their physical ______
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Fluorescence resonance energy transfer (FRET) is distance sensitive - if the distance is less than 10 nanometers, ______ happens
Fluorescence resonance energy transfer (FRET) is distance sensitive - if the distance is less than 10 nanometers, ______ happens
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Co-immunoprecipitation is an extension of IP that is based on the potential of IP reactions to capture and purify the primary target, the ______
Co-immunoprecipitation is an extension of IP that is based on the potential of IP reactions to capture and purify the primary target, the ______
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In Co-IP, complexes of two (or more) proteins are isolated using a procedure similar to the IP ______
In Co-IP, complexes of two (or more) proteins are isolated using a procedure similar to the IP ______
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Co-IP is often used for the analysis of interactions of multiple proteins and their ______
Co-IP is often used for the analysis of interactions of multiple proteins and their ______
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The principle of Co-IP is the same as IP, except that the proteins associated with the antigen are also ______
The principle of Co-IP is the same as IP, except that the proteins associated with the antigen are also ______
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A protein complex is isolated by Co-IP using an antibody for one of the components in the ______
A protein complex is isolated by Co-IP using an antibody for one of the components in the ______
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The choice of antibody is critical for successful Co-IP. The antibody must bind to the surface of the ______
The choice of antibody is critical for successful Co-IP. The antibody must bind to the surface of the ______
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