Yeast Two-Hybrid System Concepts

The video tutorial explains the yeast two-hybrid system, a method used to study protein-protein interactions. It is based on the gal4 transcription factor, which requires two domains to function. The system uses two plasmids, each containing a domain of gal4 and a protein of interest. When transformed into yeast cells, if the proteins interact, the domains come together to activate a reporter gene, indicating interaction. Detection can be through fluorescence or colorimetric assays.

What is the primary purpose of the yeast two-hybrid system?

• To study DNA replication

• To analyze protein-protein interactions

• To examine lipid metabolism

• To investigate RNA synthesis

Which transcription factor is the yeast two-hybrid system based on?

• NF-kB

• TATA

• GAL4

• SP1

In the yeast two-hybrid system, what happens if the GAL4 domains are separated?

• Transcription of the downstream gene is activated

• The plasmid is degraded

• Transcription of the downstream gene is inhibited

• The yeast cell dies

What is the role of plasmids in the yeast two-hybrid system?

• To synthesize RNA

• To carry genes for GAL4 domains and proteins of interest

• To degrade proteins

• To replicate DNA

How are the genes for proteins of interest incorporated into the yeast two-hybrid system?

• By using viral vectors

• By fusing them with yeast DNA

• By ligating them next to GAL4 domains in plasmids

• By direct injection into yeast cells

What is the significance of transforming plasmids into yeast cells in this system?

• To enable protein synthesis

• To produce energy

• To allow the expression of GAL4 domains and proteins of interest

• To initiate cell division

What occurs if protein 1 and protein 2 interact in the yeast two-hybrid system?

• The proteins are degraded

• The plasmid is expelled from the cell

• The yeast cell undergoes apoptosis

• A functional transcription factor is formed

What type of gene is typically used as a reporter in the yeast two-hybrid system?

• A gene coding for a fluorescent or colorimetric protein

• A gene coding for a structural protein

• A gene coding for a transcription factor

• A gene coding for a metabolic enzyme

What happens if protein 1 and protein 2 do not interact in the yeast two-hybrid system?

• The yeast cell divides rapidly

• The downstream gene is not transcribed

• The plasmid is replicated

• The downstream gene is transcribed

Which of the following is a possible outcome of a successful protein interaction in the yeast two-hybrid system?

• Inhibition of cell growth

• Activation of a reporter gene

• Degradation of the plasmid

• Mutation of the yeast genome