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- 1. A monogenic disease is a disease caused by a mutation in a single gene. For instance, sickle-cell anemia is caused by a mutation in the HBB gene, which codes for the B- globin chain of hemoglobin. The beginning of HBB is shown here: 5'-ATGGTGCACCTGACTCCTGAGGAGAAGTCTGCCGTTACT...-3' A. Translate this HBB sequence into an amino acid sequence. B. In terms of amino acids, what is the result of the sickle cell mutation, wherein the bolded red A is changed to a T? This single mutation causes hemoglobin to aggregate, causing red blood cells to deform into a sickle-like shape rather than the normal “biconcave disk" shape. C. What would happen if the bolded blue A were mutated to at T? (This is hypothetical; it's not a mutation found in sickle-cell disease.)3). Consider the four mutations (i-iv) described below: i. One of the mutations causing cystic fibrosis in humans is a deletion of three nucleotides that eliminates a phenylalanine at position 508 of the CFTR protein (D508). Normally, CFTR protein is localized to the plasma membrane, where it functions as a chloride ion channel. D508 CFTR is misfolded and all of it is degraded without ever reaching the cell surface. ii. The yeast transcription factor Gal4p contains a DNA-binding domain and a transcriptional activation domain. An allele with a deletion the gene portion encoding the activation domain encodes a truncated Gal4p containing only the DNA-binding domain. It binds to Gal4p target genes at appropriate binding sites in their upstream regulatory regions, but does not activate their transcription. In cells with both wild type and mutant forms of Gal4p, the truncated Gal4p binds more efficiently to target DNA sequences than wild type. iii. Mutations in the acid maltase gene in…1. A variety of organic chelating ligands have been synthesized to tightly coordinate radioactive metal cations to identify and treat malig- nancies by coupling the metal complex to a polypeptide linker attached to a monoclonal antibody that binds specifically to a cell surface re- ceptor. One such metal complex is illustrated in the diagram on the right. The radioactive Cu-64 cation is tightly coordinated by the che- lating ligand that is, in turn, conjugated to a peptide linker attached to a monoclonal antibody. (The antibody is not shown in this diagram.) When coupled to a specifically designed monoclonal antibody, the complex binds specifically to somatostatin receptors that are ex- pressed on the surface of neuroendocrine tumors. Subsequently the entire complex with the receptor is internalized, i.e., passed into the cytoplasm, where the radioactive metal cation kills the malignant cell. CH3 HN NHNH HN NHNH HN НО НО NH HN HN HO S-S NH (a) dues in the polypeptide. Place the label…
- 1. Using the data available describe the phenotype that you observe when each of these three genes is knocked down using RNAi. Remember to compare the worms on control RNAi plates with worms on each of the RNAi knockdown plates. In each case state how do these phenotypes relate to the function of the gene. (gene name for F27C1.8 is dpy-5,for T19E7.2 is skn-1) 2. Describe further experiments that you could do that would confirm your findings.1. "Interferons (IFNs)‐α and ‐β are expressed in response to a virus infection and are released from the cell in which they are produced. IFNs induce an antiviral state in other neighboring cells. a. Which cellular process is inactivated when IFN‐treated cells are infected with a virus? b. One arm of the IFN‐induced antiviral state is the synthesis of 2′,5′‐oligoA in response to viral infection. In one sentence or a simple diagram, what is the effect of this on the cell? c. Another arm of the IFN‐induced antiviral state is activation of the protein kinase in response to viral infection. In one sentence or a simple diagram, what is the effect of this on the cell? d. All cells contain the genes for IFNs. IFN synthesis is stimulated by virus infection. Would you expect a cell that has been treated with IFN to synthesize IFN in response to a viral infection? Explain your answer."2. For a fictional human tissue, it is hypothesised that cancer stem cells have the following expression pattern of cell surface markers (CD44+/CD24+/ESA+). Flow cytometry was used to isolate this population of cells. COMM CD24 ESA 1% Tumour volume (cm³) a) In the flow cytometry experiment, explain how to determine what percentage of the total cell population are the putative cancer stem cells. Also, briefly describe two controls that should be included in the experimental design. (30%) 3.01 2.5- +602 In a follow-up experiment, sorted and unsorted cells were injected subcutaneously into immunodeficient mice. The following plot depicts the results from the experiment. 2.0- 1.5- 1.0- CD24 0.5- ESA 37% • Unsorted CD44+/CD24+/ESA+ CD44+/CD24-/ESA- Time (weeks) 10 12
- 6_c. In cancerous cells, CpG islands are: where intercalating agents are found demethylated methylated.3. Coronaviruses express a nucleocapsid protein that is needed for propagation, transcription, and assembly of the virus. The nucleocapsid protein must be phosphorylated by a kinase in the host cell to carry out these functions. One such kinase that has been recently reported is glycogen synthase kinase 3 or GSK-3. The following is the 10-letter sequence of the nucleocapsid protein that is recognized and phosphorylated by GSK-3: SSRGTSPARM. Note: pk. N-terminus = 9.3; pk. R = 12.5; pK. T= 13; pK. S= 13; pK. C-terminus = 4.3 a) What is the sequence of the peptide using the three-letter amino acid abbreviations? b) Draw the chemical structure of the peptide when it is at pH 8. Assign charges and label the peptide bonds. c) What is the pl of the peptide? Do not use an online resource to calculate this value. Show your work to receive credit.You are investigating the activity of different cadherins in cells. You use recombinant DNA technology to express GFP-E-cadherin (green) in some L-cells (which do not normally contain cadherin) and YFP-P-cadherin (yellow) in other L-cells. Explain results you expect in three cultures containing: (1) only cells expressing GFP-E-cadherin, (2) only cells expressing YFP-P-cadherin, and (3) a mixture of GFP-E-cadherin-expressing and YFP-P-cadherin-expressing cells. Also predict how (a) knocking down the ability of the cells to synthesize fibronectin (FN) or (b) adding EGTA, which depletes free Ca2+ concentration in the culture medium, might affect the results.
- 7. Consider the 3 mechanisms of HGT in prokaryotes and identify 1) which one leads to the presence of a spacer in the CRISPR locus of a bacterial chromosome and 2) describe how this HGT mechanism occurs. Hint: in 2) the question asks to describe the HGT mechanism, not how the spacer is incorporated into the bacterial chromosome by Cas proteins. The HGT mechanism that leads to a spacer in a bacterial chromosome is During this mechanism/20. In class, we discussed diffferent types of genetic change that can cause a normal gene (proto- oncogene) to become a cancer-causing gene (oncogene), Which of the following would not be a cause? A) translocation or transposition (movement of DNA within the same genome) B) gene ampliffication (increased number of copies of a given gene) C) epigenetic change D) point mutation that changes the gene's product E) loss of telomeres during DNA replication ancer?2) How does the HAEIII enzyme discriminate between the C-G polymorphism in the TAS2R38 gene?