You are a researcher who has just discovered a new protein in a fungus. To help
ID: 221027 • Letter: Y
Question
You are a researcher who has just discovered a new protein in a fungus. To help determine the protein's function, you want to identify the other proteins in the fungal cell with which your protein interacts. How do you design a yeast two-hybrid experiment to address this problem?
A) Express the protein in bacterial strain 1 as a fusion protein with the DNA-binding domain of Gal4p. Using bacterial strain 2, make a library representing all fungal proteins expressed as fusion proteins with the Gal4p activation domain. Encourage conjugation between strain 1 and strain 2, and look for colonies that express a reporter gene. These colonies will generally arise from cells containing fusion protein that interacts with your target protein. B) Express the protein in yeast strain 1 as a fusion protein with the ribosomal-binding domain of Gal4p. Using yeast strain 2, make a library representing all fungal proteins expressed as fusion proteins with the Gal4p activation domain. Mate strain 1 with the strain 2 library, and look for colonies that express a reporter gene. These colonies will generally arise from mated cells containing fusion protein that interacts with your target protein. C) Express the protein in yeast strain 1 as a fusion protein with the DNA-binding domain of Gal4p. Using yeast strain 2, make a library representing all fungal proteins expressed as fusion proteins with the Gal4p activation domain. Mate strain 1 with the strain 2 library, and look for colonies that express a reporter gene. These colonies will generally arise from mated cells containing fusion protein that interacts with your target protein. D) Express the protein in yeast strain 1 as a fusion protein with the DNA-binding domain of Gal4p. Using yeast strain 2, make a library representing all fungal proteins expressed as fusion proteins with the Gal4p activation domain. Mate strain 1 with the strain 2 library, and look for colonies that express the target gene, which creates either a color or another selective advantage. These colonies will generally arise from mated cells containing fusion protein that interacts with your target protein.Explanation / Answer
Answer: Option D is correct.
Explanation:
In yeast two-hybrid assay,
Bait protein is fused to the GAL4 DNA binding domain.
The prey proteins are fused to the GAL4 activation domain.
Both these domains are brought together only when the bait and prey proteins interact with each other.
The reconstitution of the AD+BD results in the transcription of a visible marker gene such as galactosidase or a nutritional marker such as Ade.
Related Questions
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.