If I transform an ade- (adenine minus) ura- (uracil minus) E. coli strain with wild type...
If I transform an ade- (adenine minus) ura- (uracil minus) E. coli strain with wild type E. coli chromosomal DNA. How should I select for ade+ transformants?
Spread the transformation solution on a minimal medium + adenine
Spread the transformation solution on a minimal medium + uracil
Spread the transformation solution on a minimal medium + streptomycin
Spread the transformation solution on a minimal medium + adenine + uracil
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If I transform an ade- (adenine minus) ura- (uracil minus) E.
coli strain with wild type...
You have a strain of E. coli which is ade- (adenine minus, meaning unable to make...
You have a strain of E. coli which is ade- (adenine minus, meaning unable to make adenine on its own). In the biosynthesis of adenine, there are five steps as follows: In the biosynthetic pathway shown above, the ‘precursor’ is the starting material. Enzyme 1 converts this precursor to compound 1 (intermediate 1), enzyme 2 then acts on compound 1 to give rise to compound 2 (intermediate 2), so on and so forth, until enzyme 5 gives rise to the...
1. An auxotrophic E. coli strain requires adenine to grow because of a mutation in a...
1. An auxotrophic E. coli strain requires adenine to grow because of a mutation in a gene for an adenine synthesis enzyme. The following shows part of the wild-type and mutant alleles of the gene, including the start codon. The bottom strand is the template for transcription. How does this mutation affects the enzyme? oder wild- type allele ode mutant allele S'...TTATGGGCAAGATCCCA... 3 3...AATACCCGTTCTAGGGT... 5 ... TTATGGGCTAGATCCCA...3 3...AATACCCGATCTAGGGT...5
You are conducting an experiment using wild-type E. coli and you find that the beta-galactosidase activity...
You are conducting an experiment using wild-type E. coli and you find that the beta-galactosidase activity of a culture grown in glycerol minimal medium is the same as a culture grown in lactose minimal medium. Is this expected? True or false: Glycerol is another possible carbon source for E. coli and it is not either glucose or lactose.
III. You isolate a new species of bacteria that only grows in the Antarctic. A wild...
III. You isolate a new species of bacteria that only grows in the Antarctic. A wild type strain of this bacteria can synthesize its own alanine (alat) and its own tryptophan trp' ), both of which are essential amino acids. A mutant strain of the same bacterium cannot synthesize either of those two amino acids (ala trp'). You are interested in the degree of linkage between the ala and the trp genes in this new bacterial species. Fortunately, this species...
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...
3. Consider a hypothetical strain of E. coli (strain 401) that contains two mutations that affect...
3. Consider a hypothetical strain of E. coli (strain 401) that contains two mutations that affect tryptophan biosynthesis. The first mutation is a single nucleotide substitution that converts the second tandem Trp codon in region 1 of the Trp leader sequence into a stop codon (see slides 2-7 of Lecture 24 on iLearn) trp structural genes trpE trpD trpC trpB trpA PO Trp Leader Met-Lys-Ala-lle-Phe-Val-Leu-Lys-Gly-Trp-Trp-Arg-Thr Ser- Stop Assume that strain 401 also contains a mutation in the gene for the...
You have found a strain of microorganism, Schizosaccharomyces spp that is very sensitive to mutagenic compounds....
You have found a strain of microorganism, Schizosaccharomyces spp that is very sensitive to mutagenic compounds. When grown in the presence of known mutagens this organism produces many mutant progeny. This microbe, initially isolated from beer-stained carpeting from Zach’s apartment, is obviously deficient in repairing damage to DNA. You perform a series of experiments to determine which repair system is defective. A. You first take two identical cultures and expose them to 30 minutes of irradiation. You place one of...
There are two different strains of E. coli. One type is a wild type and the other is a lac operon mutant. The lac operon wild type has the genotype: I+P+O+Z+Y+A+ What is a silent mutation? What w...
There are two different strains of E. coli. One type is a wild type and the other is a lac operon mutant. The lac operon wild type has the genotype: I+P+O+Z+Y+A+ What is a silent mutation? What would be the result for the lac operon genes?
2 What is the minimum genotype of a recombinant cell that grew on minimal media supplemented...
2 What is the minimum genotype of a recombinant cell that grew on minimal media supplemented with arginine, methionine and the antibiotic tetracycline but lacking the essential amino acid isoleucine 3 How would you determine if your recombinant cells had also acquired the tryptophan gene from the Hart Complete the following table indicating which of the organisms (F and/or Hfr used in this experiment would be expected to grow on the given media. Unless specified, MMD is just minimal medium...
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...
1. An auxotrophic E. coli strain requires adenine to grow because of a mutation in a gene for an adenine synthesis enzyme. The following shows part of the wild-type and mutant alleles of the gene, including the start codon. The bottom strand is the template for transcription. How does this mutation affects the enzyme? oder wild- type allele ode mutant allele S'...TTATGGGCAAGATCCCA... 3 3...AATACCCGTTCTAGGGT... 5 ... TTATGGGCTAGATCCCA...3 3...AATACCCGATCTAGGGT...5
III. You isolate a new species of bacteria that only grows in the Antarctic. A wild type strain of this bacteria can synthesize its own alanine (alat) and its own tryptophan trp' ), both of which are essential amino acids. A mutant strain of the same bacterium cannot synthesize either of those two amino acids (ala trp'). You are interested in the degree of linkage between the ala and the trp genes in this new bacterial species. Fortunately, this species...
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...
3. Consider a hypothetical strain of E. coli (strain 401) that contains two mutations that affect tryptophan biosynthesis. The first mutation is a single nucleotide substitution that converts the second tandem Trp codon in region 1 of the Trp leader sequence into a stop codon (see slides 2-7 of Lecture 24 on iLearn) trp structural genes trpE trpD trpC trpB trpA PO Trp Leader Met-Lys-Ala-lle-Phe-Val-Leu-Lys-Gly-Trp-Trp-Arg-Thr Ser- Stop Assume that strain 401 also contains a mutation in the gene for the...
2 What is the minimum genotype of a recombinant cell that grew on minimal media supplemented with arginine, methionine and the antibiotic tetracycline but lacking the essential amino acid isoleucine 3 How would you determine if your recombinant cells had also acquired the tryptophan gene from the Hart Complete the following table indicating which of the organisms (F and/or Hfr used in this experiment would be expected to grow on the given media. Unless specified, MMD is just minimal medium...
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...
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