Coursework Question:
Answer all three parts of the question. Use diagrams and flow charts as well as concise free text responses and detailed figure legends as required. Text in figure legends will be included in the overall word count. There are 5 genes that are known to be commonly associated with a rare human disease (disease X). Not all patients with disease X have mutations in one or more of these genes, and patients often exhibit differences in disease severity and age of onset.
Comparative genomics shows that the 5 disease associated genes are conserved in all eukaryotic cells, although deletion of these genes induces different phenotypes in different model organisms (Figure 1).
1). With the aid of a diagram briefly explain the principles and processes by which it may be possible to identify novel human genes associated with disease X. In your answer, you should consider the fact that protein complexes are often conserved through evolution, however, although orthologues may be highly conserved, they may perform different functions in different species. Indicative length approx. 700 words
2). Using either C. elegans or Drosophila, design an experimental strategy, involving a classic forward genetic approach, to identify additional genes that have the ability to suppress disease X, or act as new therapeutic targets to modify the severity of disease X in humans. Indicative length approx. 700 words
3). Describe the principle of the yeast two hybrid assay and explain why different results may be seen on (-) His or (-) Ade selection plates. Figure 2 shows the results of a targeted yeast two-hybrid screen. Considering that the relative intensity of grey reflects the extent of growth on (-) Ade and (-) His selective plates (Darker = more growth), order the novel interaction partners by dissociation constant (Kd) values that you may expect if these interactions were retested by Fluorescence Cross Correlation Spectroscopy (FCCS) in human cells, and briefly explain your reasoning. Indicative length approx. 600 words
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