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Homework Answers
His3 is an important enzyme for histidine biosynthesis, therefore, it is required for the survival of yeast in His- media (histidine auxotrophs). CYP*-His3 fusion protein will help yeast survival in His- media. Wild type yeast will degrade CYP* protein and also CYP*-His3 fusion protein, which means it will not survive in His- media. Similarly rkr1 mutant degraded CYP* protein and also CYP*-His3 fusion protein, and not survive in His- media. HRD1 mutant did not degrade CYP* protein and CYP*-His3 fusion protein which means it is an important enzyme in CYP*- degradation. DOA10 mutant follow similar pattern as HRD1 but with weaker intensity.
Based on the data provided in 5 the western blot will be as follow. Note Pgk1 is housekeeping reference gene which shows equal protein is loaded in all samples.
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please help with part A and B. 2. A yeast geneticist isolates two different haploid mutant...
please help with part A and B.
2. A yeast geneticist isolates two different haploid mutant yeast strains, Strain A and Strain B, which cannot grow unless the amino acid leucine is added to the growth media. Wild-type yeast strains can make their own leucine and do not require that it be added to the growth media. The geneticist discovers that each mutant yeast strain contains a single recessive mutation that leads to the observed leucine-requiring phenotype. When she crosses...
6. (20 points) You have created a green fluorescent protein (GFP) fusion to a protein that...
6. (20 points) You have created a green fluorescent protein (GFP) fusion to a protein that is normally secreted from yeast cells. Because you are interested in studying protein and vesicle transport, you obtain three mutant yeast strains, each defective in some aspect of the protein secretory process. Being a good scientist, you of course also obtain a wild-type control strain. You decide to examine the fate of your GFP fusion protein in these various yeast strains and engineer the...
Mutation Cremoves the signal sequence from prolon 3 Mutation D prevents the fusion of all vesicles...
Mutation Cremoves the signal sequence from prolon 3 Mutation D prevents the fusion of all vesicles to the cell membrane 2. You are studying a transmembrane cell surface protein called Hip in yeast cells. You generate a wild-type Hip-GFP fusion protein and 2 mutant proteins as shown below: M Wild-type protein-GFP Hip GFP Stretch of 6 hydrophobic amino acids Stretch of 18 hydrophobic amino GFP Mutant 1-GFP N- Mutant 2-GFP N- You examine yeast cells expressing the above proteins separately,...
Q 3: You are trying to identify the peroxisome-targeting sequence in the thiolase enzyme in yeast....
Q 3: You are trying to identify the peroxisome-targeting sequence in the thiolase enzyme in yeast. The thiolase enzyme normally resides in the peroxisome and therefore must contain amino acid sequences that are used to target the enzyme for import into the peroxisome. To identify the targeting sequences, you create a set of hybrid genes that encode fusion proteins containing part of the thiolase protein fused to another protein, histidinol dehydrogenase (HDH). HDH is a cytosolic enzyme required for the...
QUESTION 7 Imagine you discovered a strain of E. coli lacking a gene necessary to synthesize...
QUESTION 7 Imagine you discovered a strain of E. coli lacking a gene necessary to synthesize the amino acid histidine (they are therefore his). You attempt to rescue the cells by transforming them DNA from a wild-type his "donor" strain, and you plate your transformed sample and all controls on plates lacking histidine (-His) as well as plates containing histidine (+His). Shown below are the results of your transformation. + indicates there was growth, and - indicates there was no...
Genetics problem: I am looking for help with (e) but I have provided (d) for the...
Genetics problem: I am looking for help with (e) but I have
provided (d) for the added input it has on the problem set. Thank
you to whomever is able to answer the problem.
(d) (2 pts) F' merodiploid strains are constructed for the region of the chromosome that includes the histidine biosynthetic operon. Below are the results of pairwise complementation tests: a"+" indicates growth without added histidine: F' His Donor His mutant# 10 4. 2 his Growth on minimal...
. Gleevec (Imatinib) inhibits protein kinase BCR-ABL, which is constitutively active in patients with Chronic Myelogenous...
. Gleevec (Imatinib) inhibits protein kinase BCR-ABL, which is
constitutively active in patients with Chronic Myelogenous Leukemia
(CML). The structure of Gleevec is shown below:
a) Based on our discussion in lecture and the structure above,
explain how the “lead” compound structure for Gleevec was designed
and how it led to the development a compound that could bind ABL
with higher affinity than ATP.
b) Using a broad kinase inhibition assay, you discover that
Gleevec inhibits another tyrosine kinase called...
QUESTION 1 Imagine you are preparing an industrial enzyme assay. The protocol says to add 20...
QUESTION 1 Imagine you are preparing an industrial enzyme assay. The protocol says to add 20 mL of 100 mM NaCl to your assay, and the final volume of the assay is 100 ml. What is the working concentration of NaCl? The number of significant digits you include in your answer doesn't matter, but it must be in units of mM. QUESTION 2 Imagine you are preparing an enzyme assay that requires a working concentration of 10 mM NaCl. If...
Which technique would you use if you wanted to < Identify all the regions of open...
Which technique would you use if you wanted to < Identify all the regions of open chromatin in a particular cell type? Perform a paternity test? Amplify a specific sequence of DNA? [Choose] [Choose] Promoter fusion DNAse-seq Scanning mutagenesis Western blot RNAseq DNA fingerprinting Northern blot Reporter Assay DNAse footprinting ChIP-PCR EMSA Southern blot DNAse protection assay PCR BLAST search Protein fusion ChIP-seq [Choose Compare the global level of RNA expression between two samples? Detect presence of a specific protein...
This question will help you predict the outcome of the FOA assay in week 11. Prior...
This question will help you predict the outcome of the FOA assay in week 11. Prior knowledge: (1) The genotype of the yeast reporter strain AGY75 is "trp1; ura3; his 3; msh2A; PSH44". It carries the mutated/non-functional alleles for gene Trp1; Ura3; His3; a deletion that removed MSH2; and the PSH44 plasmid. (2) PMSH2 plasmid carries gene AmpR, His 3, MSH2 (3) pSH44 plasmid carries gene GT16.5-Ura3, Trp1 (4) SD/-His, -Trp, -Leu, -Thr, +FOA plate: SD medium containing FOA, and...
please help with part A and B.
2. A yeast geneticist isolates two different haploid mutant yeast strains, Strain A and Strain B, which cannot grow unless the amino acid leucine is added to the growth media. Wild-type yeast strains can make their own leucine and do not require that it be added to the growth media. The geneticist discovers that each mutant yeast strain contains a single recessive mutation that leads to the observed leucine-requiring phenotype. When she crosses...
6. (20 points) You have created a green fluorescent protein (GFP) fusion to a protein that is normally secreted from yeast cells. Because you are interested in studying protein and vesicle transport, you obtain three mutant yeast strains, each defective in some aspect of the protein secretory process. Being a good scientist, you of course also obtain a wild-type control strain. You decide to examine the fate of your GFP fusion protein in these various yeast strains and engineer the...
Mutation Cremoves the signal sequence from prolon 3 Mutation D prevents the fusion of all vesicles to the cell membrane 2. You are studying a transmembrane cell surface protein called Hip in yeast cells. You generate a wild-type Hip-GFP fusion protein and 2 mutant proteins as shown below: M Wild-type protein-GFP Hip GFP Stretch of 6 hydrophobic amino acids Stretch of 18 hydrophobic amino GFP Mutant 1-GFP N- Mutant 2-GFP N- You examine yeast cells expressing the above proteins separately,...
Q 3: You are trying to identify the peroxisome-targeting sequence in the thiolase enzyme in yeast. The thiolase enzyme normally resides in the peroxisome and therefore must contain amino acid sequences that are used to target the enzyme for import into the peroxisome. To identify the targeting sequences, you create a set of hybrid genes that encode fusion proteins containing part of the thiolase protein fused to another protein, histidinol dehydrogenase (HDH). HDH is a cytosolic enzyme required for the...
QUESTION 7 Imagine you discovered a strain of E. coli lacking a gene necessary to synthesize the amino acid histidine (they are therefore his). You attempt to rescue the cells by transforming them DNA from a wild-type his "donor" strain, and you plate your transformed sample and all controls on plates lacking histidine (-His) as well as plates containing histidine (+His). Shown below are the results of your transformation. + indicates there was growth, and - indicates there was no...
Genetics problem: I am looking for help with (e) but I have
provided (d) for the added input it has on the problem set. Thank
you to whomever is able to answer the problem.
(d) (2 pts) F' merodiploid strains are constructed for the region of the chromosome that includes the histidine biosynthetic operon. Below are the results of pairwise complementation tests: a"+" indicates growth without added histidine: F' His Donor His mutant# 10 4. 2 his Growth on minimal...
. Gleevec (Imatinib) inhibits protein kinase BCR-ABL, which is
constitutively active in patients with Chronic Myelogenous Leukemia
(CML). The structure of Gleevec is shown below:
a) Based on our discussion in lecture and the structure above,
explain how the “lead” compound structure for Gleevec was designed
and how it led to the development a compound that could bind ABL
with higher affinity than ATP.
b) Using a broad kinase inhibition assay, you discover that
Gleevec inhibits another tyrosine kinase called...
QUESTION 1 Imagine you are preparing an industrial enzyme assay. The protocol says to add 20 mL of 100 mM NaCl to your assay, and the final volume of the assay is 100 ml. What is the working concentration of NaCl? The number of significant digits you include in your answer doesn't matter, but it must be in units of mM. QUESTION 2 Imagine you are preparing an enzyme assay that requires a working concentration of 10 mM NaCl. If...
Which technique would you use if you wanted to < Identify all the regions of open chromatin in a particular cell type? Perform a paternity test? Amplify a specific sequence of DNA? [Choose] [Choose] Promoter fusion DNAse-seq Scanning mutagenesis Western blot RNAseq DNA fingerprinting Northern blot Reporter Assay DNAse footprinting ChIP-PCR EMSA Southern blot DNAse protection assay PCR BLAST search Protein fusion ChIP-seq [Choose Compare the global level of RNA expression between two samples? Detect presence of a specific protein...
This question will help you predict the outcome of the FOA assay in week 11. Prior knowledge: (1) The genotype of the yeast reporter strain AGY75 is "trp1; ura3; his 3; msh2A; PSH44". It carries the mutated/non-functional alleles for gene Trp1; Ura3; His3; a deletion that removed MSH2; and the PSH44 plasmid. (2) PMSH2 plasmid carries gene AmpR, His 3, MSH2 (3) pSH44 plasmid carries gene GT16.5-Ura3, Trp1 (4) SD/-His, -Trp, -Leu, -Thr, +FOA plate: SD medium containing FOA, and...
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