The two-hybrid method facilitates the discovery of protein–protei...

Question

The two-hybrid method facilitates the discovery of protein–protein interactions. How does this technique work? Can you think of reasons for obtaining a false-positive result, that is, where the proteins encoded by two clones interact in the two-hybrid system but do not interact in the organism in which they naturally occur? Can you think of reasons you might obtain a false-negative result, in which the two proteins interact in vivo but fail to interact in the two-hybrid system?

Accepted Answer

Hello, everyone and welcome to today's video. So the two hybrid method is a powerful technique used to study protein protein interactions in living cells. In this method, two hybrid domains, a DNA binding domain which is DB D an activation domain which is ad are used to detect protein protein interactions. The DB D or the domain DNA binding domain is typically fused to a protein of interest referred to as the bait. Well, the ad or the activation domain is used to is fused to a second protein of interest is referred to as the prey. So we have a bait and a prey. When the, when the DNA binding domain and the activation domain interacts, they activate which of the following components. So as answer choice A, we have promoter region as answer choice B, we have control vectors as answer choice C we have the regulator gene and as answer choice D, we have transcription factors. So as a question states when the DNA binding domain and the activation domain interact, they're going to do it in a two hybrid method and they're going to bring the debate and they pray proteins into a very close proximity, which is going to result in the activation of a promoter region located off string of the reporter gene. Now, this promoter region is going to contain binding sites for the DNA binding domain. And when the DNA binding domain and the activation domain interact, the DNA binding domain is is brought to close proximity to the promoter which activates the transcription of the reporter gene. But when these two interact, it is the promoter region that is activated first. So the correct answer for this question is going to be a promoter region. I really hope it's helped and I hope to see you on the next video.

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