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GQ13
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- 1. Using the data available Using the data available on Moodle 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. 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. 2. Describe further experiments that you could do that would confirm your findings.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.)4. Different populations of mouse L-cells that lack cadherins were transfected with either E- cadherin or P-cadherin. If cells expressing E-cad are mixed with those expressing P-cad, they segregate into distinct balls of cells (A in figure). However, if cells expressing different levels of the same cadherin are mixed, they form a single ball, with the high expressing cells in the center B in the figure). (A) E-cadherin SORTING OUT P-cadherin (B) low P-cadherin SORTING OUT high P-cadherin Why do the cells expressing different levels of cadherin segregate as in (B) in the figure, and why not into two separate balls, or one ball with a different organization (randomly mixed, or high expressors on the outside)?
- 12. a. You want to create a genetic construct that will express GFP in Drosophila. In addition to the GFPcoding sequence, what DNA element(s) must youinclude in order to express this protein in flies if theconstruct were integrated into the Drosophila genome? Where should such DNA element(s) be located? How would you ensure that GFP is expressedonly in certain tissues of the fly, such as the wing?b. Suppose you insert the GFP coding region plus allof the DNA elements required by the answer to part(a)—except the enhancer—between inverted repeatsfound at the ends of a particular transposable element.Because all of the DNA sequences located betweenthese inverted repeats can move from place to placein the Drosophila genome, you can generate manydifferent fly strains, each with the construct integrated at a different genomic location. You now examine animals from each strain for GFPfluorescence. Animals from different strains showdifferent patterns: some glow green in the eyes,others in…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.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.
- 4e. You also study the expression of 3 different mutants for this gene. For each mutant answer the following: Does this mutation change the sequence of the protein produced? Why or why not? If it does change the sequence of protein be sure to write out the new sequence. If it does not change the protein sequence, what effect (if any) would you expect it to have on expression of the gene? 1 20 ORI 40 60 5'...TTCGAGCTCTCGTCGTCGAGATACGCGATGATATTACTGGTAATATGGGGATGCACTATC...3' 3' ...AAGCTCGAGAGCAGCAGCTCTATGCGCTACTATAATGACCATTATACCCCTACGTGATAG...5' * promoter2) Review the literature and identify the key players in the various mutants of the spike protein.4e. You also study the expression of 3 different mutants for this gene. For each mutant answer the following: Does this mutation change the sequence of the protein produced? Why or why not? If it does change the sequence of protein be sure to write out the new sequence. If it does not change the protein sequence, what effect (if any) would you expect it to have on expression of the gene? 1 20 ORI 40 60 5'..TTCGAGCTCTCGTCGTCGAGATACGCGATGATATTACTGGTAATATGGGGATGCACTATC...3’ 3'...AAGCTCGAGAGCAGCAGCTCTATGCGCTACTATAATGACCATTATACCCCTACGTGATAG...5’ promoter i. Mutant A has a single base pair substitution with the T/A being replaced with C/G base pair at position 35 (position denoted by the * in the sequence above). ii. Mutant B has a 2 G/C pairs inserted between position 19 and 20 (position denoted by the ^ in the sequence above).
- 2. a. You want to create a genetic construct that will express GFP in Drosophila. In addition to the GFPcoding sequence, what DNA element(s) must youinclude in order to express this protein in flies if theconstruct were integrated into the Drosophila genome? Where should such DNA element(s) be located? How would you ensure that GFP is expressedonly in certain tissues of the fly, such as the wing?b. Suppose you insert the GFP coding region plus allof the DNA elements required by the answer to part(a)—except the enhancer—between inverted repeatsfound at the ends of a particular transposable element.Because all of the DNA sequences located betweenthese inverted repeats can move from place to placein the Drosophila genome, you can generate manydifferent fly strains, each with the construct integrated at a different genomic location. You now examine animals from each strain for GFPfluorescence. Animals from different strains showdifferent patterns: some glow green in the eyes,others in…32. A 23-year-old man comes to the physician because of a 1-month history of weakness and muscle pain after vigorous exercise. His mother, sister, and grandmother have a similar condition; however, his father's sister and her children are unaffected. A muscle biopsy specimen shows ragged red fibers. This condition most likely results from a mutation in a gene located in which of the following labeled structures in the photomicrograph? 00 O O A) B) E) B A E D C AR1. How is PKD inherited? What gene is responsible for the expression of PK enzyme ?