The diagram below represents the structure of the Lucky gene, which is expressed in every cell...
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The diagram below represents the structure of the Lucky gene, which is expressed in every cell in the fruit fly. Gray boxes show exons; box sizes are not to scale. Lucky is alternatively spliced, with exon 2 skipped in all cells in the head. Assume that poly-A polymerase adds 200 nucleotides to each transcript. -40 Promoter 5+1 Translation Start Site (+101-+103) +200 +201 +1200 +1201 1400+1401+3400 +3401 V Probe mRNA from whole flies +3850 +3851 mRNA from head PolyA Stop Codon (+3701 +3703) site +3901 a) On the gel below, draw how the northern blot will look when mRNA is isolated from the whole fly, or only from the head, using a probe complementary to a portion of the mRNA in exon 1 (shown in the diagram above). Label the band sizes next to the gel. Transcription Termination Site b) Your labmate designs a different RNA probe for the Lucky gene, fluorescently labels it, and uses it in in situ hybridization. The probe shows fluorescence in the entire fly except for in the head. Where in the gene is the probe that your labmate designed? This year's Nobel Prize in Physiology and Medicine was awarded to Katalin Kariko and Dan Weissman for showing that chemically modified RNA nucleotides alter the immune response to foreign RNAs, paving the way to develop mRNA vaccines. One immune protein that binds foreign RNAs inside cells is RIG-I. The Kd of RIG-I and an unmodified RNA molecule ~100 nucleotides long is 200 nM (2 X 10 M); assume this is also true for COVID mRNA vaccines (which are really much longer). Assume there are about 5,000 RIG-I molecules in a single cell, and that during vaccination, 1,000 RNA molecules enter a single cell. The volume of a single cell is 1.67 X 10-15 L. Avogadro's number is -6 X 1023. For the questions below, show all your calculations. a) What is the concentration of vaccine RNA molecules inside cells? b) If unmodified RNA molecules are used in a vaccine, about 95% of all RNA molecules in the cell end up bound to RIG-I at equilibrium. What proportion of RIG-I molecules in the cell would be bound to RNA at equilibrium? c) An experiment showed that if the adenosines in an RNA molecule are converted to a modified nucleotide called m6A, they can still function in translation, but the Kd for RIG-I and modified RNA changes to 1100 nM (1.1 X 10 M). Do you expect a higher or lower proportion of RIG-I molecules to be bound to RNA vaccine molecules when m6A-modified nucleotides are used in the vaccine, as compared to when adenosines are not modified? The diagram below represents the structure of the Lucky gene, which is expressed in every cell in the fruit fly. Gray boxes show exons; box sizes are not to scale. Lucky is alternatively spliced, with exon 2 skipped in all cells in the head. Assume that poly-A polymerase adds 200 nucleotides to each transcript. -40 Promoter 5+1 Translation Start Site (+101-+103) +200 +201 +1200 +1201 1400+1401+3400 +3401 V Probe mRNA from whole flies +3850 +3851 mRNA from head PolyA Stop Codon (+3701 +3703) site +3901 a) On the gel below, draw how the northern blot will look when mRNA is isolated from the whole fly, or only from the head, using a probe complementary to a portion of the mRNA in exon 1 (shown in the diagram above). Label the band sizes next to the gel. Transcription Termination Site b) Your labmate designs a different RNA probe for the Lucky gene, fluorescently labels it, and uses it in in situ hybridization. The probe shows fluorescence in the entire fly except for in the head. Where in the gene is the probe that your labmate designed? This year's Nobel Prize in Physiology and Medicine was awarded to Katalin Kariko and Dan Weissman for showing that chemically modified RNA nucleotides alter the immune response to foreign RNAs, paving the way to develop mRNA vaccines. One immune protein that binds foreign RNAs inside cells is RIG-I. The Kd of RIG-I and an unmodified RNA molecule ~100 nucleotides long is 200 nM (2 X 10 M); assume this is also true for COVID mRNA vaccines (which are really much longer). Assume there are about 5,000 RIG-I molecules in a single cell, and that during vaccination, 1,000 RNA molecules enter a single cell. The volume of a single cell is 1.67 X 10-15 L. Avogadro's number is -6 X 1023. For the questions below, show all your calculations. a) What is the concentration of vaccine RNA molecules inside cells? b) If unmodified RNA molecules are used in a vaccine, about 95% of all RNA molecules in the cell end up bound to RIG-I at equilibrium. What proportion of RIG-I molecules in the cell would be bound to RNA at equilibrium? c) An experiment showed that if the adenosines in an RNA molecule are converted to a modified nucleotide called m6A, they can still function in translation, but the Kd for RIG-I and modified RNA changes to 1100 nM (1.1 X 10 M). Do you expect a higher or lower proportion of RIG-I molecules to be bound to RNA vaccine molecules when m6A-modified nucleotides are used in the vaccine, as compared to when adenosines are not modified?
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a On the gel below draw how the northern blot will look when mRNA is isolated from the whole fly or only from the head using a probe complementary to a portion of the mRNA in exon 1 shown in the diagr... View the full answer
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Cambridge International AS And A Level Biology
ISBN: 9781107636828
4th Edition
Authors: Mary Jones, Richard Fosbery, Jennifer Gregory, Dennis Taylor
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