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Topic: Cell signalling and disease experiment report

1. a) Using the information that you gathered from the bioinformatics analysis of the core promoter and proximal promoter elements, what is your interpretation of the 5’ promoter deletion analysis in control cells +/- insulin stimulation? (20 marks) – 700words
b) Outline the step by step changes in the insulin signalling pathway that would lead to an alteration in expression of this particular gene.(10 marks) – 400 words

c) What impact does palmitate have on the candidate gene and how does it influence the genes responsiveness to insulin? (5 marks) – 300words

d) What experiment would you conduct to confirm that it is transcription factors binding to the particular elements identified that are responsible for changes in promoter activity? (5 marks) – 300 words
e) Investigators would like to continue to look at the function of the protein product produced by this candidate gene, as a potential drug target. Briefly outline an experiment you could conduct in C2C12 cells to investigate the functionality of the protein (Hint: You will want to overexpress your protein within cells and look at particular downstream targets). (10 marks) – 500words
2. Background information on obesity and insulin resistance its link to type 2 diabetes, considering the role of saturated fatty acids (circulating fatty acid levels) in the development of insulin resistance in skeletal muscle. Consider cell signalling mechanisms (the insulin signalling pathway) in muscle and the use of AKT as a marker of insulin signalling pathway. Introduction to the Western Blotting technique for the analysis of phosphorylated proteins within cell signalling pathways, particularly AKT.4: Aim and objectives of lab (600words)

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