7. In molecular imaging research gene expressions in vivo can be visualized by means of the marker ferritin, which has the property of capturing iron. Which imaging technique is used to visualize this process? Explain.
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- 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' * promoter4e. 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).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. 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.2) Review the literature and identify the key players in the various mutants of the spike protein.1. Below is the abstract from a journal article “De novo methylation of CpG island sequences in human fibroblasts overexpressing DNA (cytosine-5-)-methyltransferase” by Vertino et al. Read it and answer the following questions. Recent studies showing a correlation between the levels of DNA (cytosine-5-)-methyltransferase (DNA MTase) enzyme activity and tumorigenicity have implicated this enzyme in the carcinogenic process. Moreover, hypermethylation of CpG island-containing promoters is associated with the inactivation of genes important to tumor initiation and progression. One proposed role for DNA MTase in tumorigenesis is therefore a direct role in the de novo methylation of these otherwise unmethylated CpG islands. In this study, we sought to determine whether increased levels of DNA MTase could directly affect CpG island methylation. A full-length cDNA for human DNA MTase driven by the cytomegalovirus promoter was constitutively expressed in human fibroblasts. Individual clones…
- 1. Below is the abstract from a journal article "De novo methylation of CpG island sequences in human fibroblasts overexpressing DNA (cytosine-5-)-methyltransferase" by Vertino et al. Read it and answer the following questions. Recent studies showing a correlation between the levels of DNA (cytosine-5-)- methyltransferase (DNA MTase) enzyme activity and tumorigenicity have implicated this enzyme in the carcinogenic process. Moreover, hypermethylation of CpG island-containing promoters is associated with the inactivation of genes important to tumor initiation and progression. One proposed role for DNA MTase in tumorigenesis is therefore a direct role in the de novo methylation of these otherwise unmethylated CpG islands. In this study, we sought to determine whether increased levels of DNA MTase could directly affect CpG island methylation. A full-length CDNA for human DNA MTase driven by the cytomegalovirus promoter was w ww constitutively expressed in human bro lasts. Individual…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)?
- 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. What is Transcriptome? What is Transcriptome analysis? How does the method yield information able to elucidate the structural apexes of the molecules under examination? What are the steps and primary methods used in Transcriptome analysis? What are some important applications of Transcriptome analysis?Please be as detailed as you could be in your explanation.13. An investigator is studying epigenetic markers in patients with retinoblastoma. It is most appropriate for this investigator to focus on which of the following enzymes? A) DNA methyltransferase B) DNA polymerase C) Helicase D) RNA polymerase E) Topoisomerase