RR15

Question 1

Explain yeast genome modification process

Answer 1

Allows us to understand contribution of individual genes to growth of organism

• Analyze selection of G-418 drug resistant genes, and allow them to reproduce - we can then analyze
  -Feed animals dsRNA, get phenotype corresponding to loss of specific function of gene corresponding to dsRNA

Question 2

What are the two domains of transcription factors?

Answer 2

• DNA binding domain
• transcriptional activation domain

Question 3

Explain proteomic analysis

Answer 3

Add specific proteins to TF domains and see which associate

• Create a bait and a prey on split proteins, see if they interact to reconsitute the transcription factor (could be important gene!)
• Can perform this onentire library - allowing us to figure out sections of protein interactions

Question 4

Explain protein fragment complementation

Answer 4

Separate protein into halves - neither will carry out their function
- attach split halves to bait and prey, they will come together to reconstitue function

Often used in GFP

Question 5

Explain steps in BioID

Answer 5

Biotin has very high specificity
- Fuse biotin ligase to protein of interest (A + BirA)
- Subject it to lysis - deactivated then isolated
- Identift the chosen ones using mass spectrometry