1
Q
Explain yeast genome modification process
A
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
2
Q
What are the two domains of transcription factors?
A
- DNA binding domain
- transcriptional activation domain
3
Q
Explain proteomic analysis
A
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
4
Q
Explain protein fragment complementation
A
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
5
Q
Explain steps in BioID
A
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
