RR15 Flashcards
How would you modify a yeast genome and why would you want to do this?
- Add disruption construct to diploid cell, causing homologous recombination.
- Select for G-418 drug resistance, allow them to reproduce.
This allows us to understand the contribution of individual gene to growth of organism.
What would happen if you fed animals dsRNA?
You would get a phenotype corresponding to loss of function of gene corresponding to dsRNA. This would help us understand the function of different genes. It was found that genes in post-embryonic aspects to be more specific.
What are the two domains of TF modularity? What is the outcome?
1) DNA binding
2) Transcriptional activation
- Use genomic tools to see what unknown function genes do in the cell
What is proteomic analyses?
When you add specific proteins to these domains, to see if they associate. For example, need protein A and B need to interact to reconstitute transcription factor.This could cause the activation of an important gene and allow the assessment of whether proteins come together.
You can apply this to an entire library, to figure out sections of protein interactions, which would then give hints to the function of a protein.
What is protein fragment complementation? Where is this used a lot?
It is when you separate proteins into halves and neither will carry out a function. You can attach split halves to bait and prey, the bait and prey will come together - reconstituting function.
It is used a lot in GFP, enhancing flurescence.
What is BioID? What are new versions of it?
It is a type of labelling to tag proteins in neighbourhood.
New versions:
APEX, APEXZ, TurboID
When would you use heterologous gene product to tag proteins?
If the proteins come close to the area of the protein of interest. You can use ofecoli enzyme.
If you found a BirA mutation that releases biotin, what could you do?
You could transfect cells with protein + BirA. This is a biotin molecule tag that makes it easier to purify.
When would you use
