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Flashcards
Technologie
Proteomics
The yeast two –hybrid assay
- Prinzip
The yeast two hybrid assay uses the fact that many eukaryotic transcriptional regulators have two parts or domains that can act independently of one another. One of them is the so-called DNA binding domain (DBD), which is responsible for binding of the transcription factor to DNA near a gene whose expression it regulates. The second is the transcriptional activation domain (TAD), which is responsible for interacting with a protein complex (not shown in the slide) that includes RNA polymerase, and that is bound near the transcriptional start site of the regulated gene. This interaction triggers the initiation of transcription of the gene. The yeast two-hybrid assay asks whether two proteins of interest (A,B) can physically interact. It requires fusing one protein to the DBD of a transcriptional regulator, and the other to the TAD of this regulator. These two ‘hybrid’ proteins are also sometimes called ’bait’ and ‘prey’, respectively. If the two proteins A and B can interact, they can bring these two domains together, which can then activate a gene’s expression. The gene used in this assay is a so-called reporter gene whose expression can be easily detected. It is merely used to ‘report’ whether gene expression occurred, and whether the two proteins A and B interacted.
Technologie
structural Proteomics
Determining protein structure
- Prinzip
-> welche Ansätze gibt es
- Applications
Technologie
Reverse Genetics
RNAi
- was ist das?
- wo wurde es entdeckt?
905
RNAi depends on a system that the organism uses to detect double-stranded RNA viruses as well as other abnormal gene expression. This mechanism was initially characterized in the fungus Neurospora, where the phenomenon is known as quelling, and in plants, where it is responsible for resistance to the spread of virus.
Menschliche Genomvariation
Mendelian diseases
- Huntignton
- ZF
- Hämophilie
=> VErerbung
- Huntignton -> Dominant
- ZF -> rezessiv
- Hämophilie -> X-Linked