Describe the principle of the yeast-two hybrid system
2) A positive result for the yeast two-hybrid analysis means:
A. two fusion proteins, one containing the Gal4p binding site domain and one containing the Gal4p activation domain, have interacted through their Gal4p domains resulting in transcription of the reporter gene.
B. two fusion proteins, one containing the Gal4p binding site domain and one containing the Gal4p activation domain, have bound together, which disrupts the ability of the reporter gene to make a blue colony.
C. two fusion proteins interact with each other and because the Gal4p binding site domain on one fusion protein binds to the reporter gene and the Gal4p activation domain of the other fusion protein interacts with RNA polymerase, the reporter gene is activated.
D. two fusion proteins interact with each other and because of the Gal4p binding site domain and the Gal4p activation domain contained in each fusion protein, the reporter genes are activated and the cells die, resulting in no colonies. E. the colonies growing are expressing the two fusion proteins and the two proteins have functioned independently.
3) What could the reporter gene encode and how would you detect reporter gene expression?
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