During the first successful gene therapy trial in which Ashanti DeSilva was treated for SCID, did the therapeutic gene delivered to Ashanti replace the defective copy of the ADA gene? Why were white blood cells chosen as the targets for the therapeutic gene?
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During the first successful gene therapy trial in which Ashanti
DeSilva was treated for SCID, did the therapeutic gene delivered
to Ashanti replace the defective copy of the ADA gene? Why
were white blood cells chosen as the targets for the therapeutic
gene?
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- In the plaque assay, exactly what makes up a single plaque?In selecting target cells to receive a transferred gene in gene therapy, what factors do you think would have to be taken into account?Draw the Western Blot that would result from undertaking the same experiment in the presence of: i) An estrogen receptor agonist ii) An estrogen receptor antagonist iii) An estrogen receptor agonist in the presence of an excess amount of antagonist iv) Estrogen in the presence of an excess amount of antagonist v) An estrogen receptor inverse agonist.
- The strain of λ phage t is cI857. That tells you that the cI DNA segment is disabled by a specific mutation. What is the exact genetic change in cI857? What specific property of the cI gene product does this mutation change, and how does this help titering for a plaque assay?You decide to use a plaque assay with a human lung epithelial cell line to compare the effectiveness of the drug. The results of the plaque assay for one strain of influenza are shown below. Based on these results, what can you conclude about the relative effectiveness of WJ379 at 1 mM compared to oseltamivir at 1 mM? Briefly explain your rationale.The Pfizer Covid-19 vaccine is one of the vaccines currently being rolled out for mass vaccination in South Africa to protect the population against SARS CoV-2, and the active compound in this vaccine is an mRNA molecule. a) The starting material for productio of this vaccine is a copy of the coronavirus spike protein gene cloned into a ppasmid. List the main experimental steps in the vaccine production process, to go from a plamid to an mRNA molecule. b) Which molecular processes in the cell will allow an injected strand of mRNA to produce anti-Covid immunity in the vaccine recipient?
- Which vector is used to replace the defective gene in gene therapy?Bacteriophage P22 was used in generalized transduction experiments to infect the Salmonella typhimurium donor strains described in the table below. The resulting phage lysates were then used to infect the S. typhimurium recipient strains listed in the table. In each cross, a phenotype was selected for one of the three genetic markers studied (str, aceA, thrA), and then replicates were performed to select the corresponding recombinants for the other two markers. The results are given in the following table: Recipient strain Selected phenotype Selected recombinants Donor strain str thrA aceA+ thrA str aceA+ strs thrA+ aceA thrA+ str aceA Str Ace+ Str ThrA ThrA+ ThrA ThrA+ Ace Ace str: gene involved in streptomycin resistance, aceA gene involved in the use of acetate as a carbon source, thrA: gene involved in the biosynthesis of threonine. Number 60 40 95 5 10 90 Determine the order of the genes and draw a genetic map showing this orderBacteriophage P22 was used in generalised transduction experiments to infect the Salmonella typhimurium donor strains described in the table below. The resulting phage lysates were then used to infect the recipient strains of S. typhimurium recipient strains listed in the table. In each cross, a phenotype was selected for one of the selected for one of the three genetic markers studied (str, aceA, thrA), and were made to select the recombinants corresponding to the other two markers. markers. The results are given in the following table: Strain I donor str thrA aceA thrA str aceA+ Strain recipient strs thrA+ aceA thrA str aceA Phenotype selected Str Ace+ Str recombinants selected ThrA ThrA ThrA ThrA Ace Ace Number 60 40 95 5 10 90 str: gene involved in streptomycin resistance, aceA: gene involved in the use of acetate as a carbon source, thrA: gene involved in threonine biosynthesis. 1) What are the selective media used in these three transduction experiments? to obtain the selected…
- What does MEGA stand for and what is this assay used to determine? How to do perform the assay?Explain how site directed mutagenesis can be accomplished using M13 bacteriophage. Using this method often results in only 5% of plaques with mutant gene. Explain how the procedure has been modified to enrich for the number of plaques with the mutant gene.a. Bacteriophage P22 was used in generalized transduction experiments to infect the Salmonella typhimurium donor strains described in the table below. The resulting phage lysates were then used to infect the S. typhimurium recipient strains listed in the table. In each cross, a phenotype was selected for one of the three genetic markers studied (str, aceA, thrA), and then replicates were performed to select the corresponding recombinants for the other two markers. The results are given in the following table: Recipient strain Selected phenotype Selected recombinants Donor strain str thrA aceAt thrA str aceA+ strs thrA+ aceA thrA+ str aceA Str Ace+ Str ThrA ThrA+ ThrA ThrA+ Ace Ace+ str: gene involved in streptomycin resistance, aceA: gene involved in the use of acetate as a carbon source, thrA: gene involved in the biosynthesis of threonine. Number 60 40 95 5 10 90 What are the selective media used in these three transduction experiments, on the one hand to obtain the selected…