Genetics: From Genes to Genomes
6th Edition
ISBN: 9781259700903
Author: Leland Hartwell Dr., Michael L. Goldberg Professor Dr., Janice Fischer, Leroy Hood Dr.
Publisher: McGraw-Hill Education
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Textbook Question
Chapter 14, Problem 15P
E. coli cells usually have only one copy of the F plasmid per cell. You have isolated a cell in which a mutation increases the copy number of F to three to four per cell. How could you distinguish between the possibility that the copy number change was due to a mutation in the F plasmid versus a mutation in a chromosomal gene?
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Students have asked these similar questions
E. coli cells usually have only one copy of the F plasmidper cell. You have isolated a cell in which a mutationincreases the copy number of F to three to four per cell.How could you distinguish between the possibility thatthe copy number change was due to a mutation in theF plasmid versus a mutation in a chromosomal gene?
Scientists modified the tumor-inducing (Ti) plasmid, a naturally occurring plant plasmid found in Agrobacterium tumefaciens, to create a tool used to introduce any gene of interest into plant cells. They created a binary system because a single Ti plasmid is too large to be easily manipulated. One part of the system is a disarmed
plasmid, and the second part is a transformation vector. The following sentences describe the function of key DNA elements in the system.
Virulence
region
Genes for
conjugative transfer
Disarmed
Ti plasmid
(T-region removed)
ori
Kan selectable marker
plant selectable marker
constitutive promoter
T-region
conjugative transfer
virulence
Kan
(bacterial
selectable
marker)
Amp selectable marker
Plant selectable marker
(e.g., herbicide resistance)
Constitutive
promoter
T-DNA left
border
Place the terms in the appropriate blanks to complete the sentences. Not all terms will be used.
3' transcriptional terminator
MCS (inserted
gene of interest)
T-region
Transformation…
You have two cell cultures, each
containing a different plasmid. The first
plasmid is ~5kbp long and contains three
5'-TCGA-3' and the other is the same size,
but only contains two 5'-TCGA-3'
sequences. You forget to label the two cell
cultures you grew and have to figure out
which one contains each plasmid. How
would you go about identifying the the
two cultures.
Chapter 14 Solutions
Genetics: From Genes to Genomes
Ch. 14 - Choose the phrase from the right column that best...Ch. 14 - The unicellular, rod-shaped bacterium E. coli is 2...Ch. 14 - Now that the sequence of the entire E. coli K12...Ch. 14 - Bacterial genomes such as that of E. coli...Ch. 14 - List at least three features of eukaryotic genomes...Ch. 14 - Describe a mechanism by which a gene could move...Ch. 14 - High salt concentrations tend to cause protein...Ch. 14 - Recently, scientists tested the possibility that...Ch. 14 - A recent metagenomic study analyzed the...Ch. 14 - Linezolid is a new type of antibiotic that...
Ch. 14 - A liquid culture of E. coli at a concentration of...Ch. 14 - Pick out the medium i, ii, iii, or iv onto which...Ch. 14 - This problem concerns Fig. 14.14, which...Ch. 14 - In two isolates one is resistant to ampicillin,...Ch. 14 - E. coli cells usually have only one copy of the F...Ch. 14 - In E. coli, the genes purC and pyrB are located...Ch. 14 - DNA sequencing of the entire H. influenzae genome...Ch. 14 - Genes encoding toxins are often located on...Ch. 14 - a. You want to perform an interrupted-mating...Ch. 14 - In Problem 19, do you think that most of the...Ch. 14 - One issue with interrupted-mating experiments such...Ch. 14 - Prob. 22PCh. 14 - Starting with an F- strain that was prototrophic...Ch. 14 - You can carry out matings between an Hfr and F...Ch. 14 - Genome sequences show that some pathogenic...Ch. 14 - Generalized and specialized transduction both...Ch. 14 - This problem highlights some useful variations of...Ch. 14 - A researcher has a Trp auxotrophic strain of E....Ch. 14 - Streptococcus parasanguis is a bacterial species...Ch. 14 - The sequence at one end of one strand of the...Ch. 14 - Scientists who study amino acid biosynthesis...Ch. 14 - Suppose that you could obtain radioactively...Ch. 14 - Prob. 34PCh. 14 - Some scientists are trying to engineer...
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Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.Similar questions
- The Molecular Cell Biology Unit of University X is working on a particular gene segment called ABC20. For a specific kind of experiment, they need to quantify the gene and require a lot of copies of the gene. The initial gel run showed a very faint band of the gene which made the quantification process difficult to proceed. Now propose a method with the help of which they can generate many copies of the gene. Here you need to know that they have the plasmid from where they can get the gene. You do not need to explain the process in detail rather answer in no more than 10 sentences on what can be done about this problem!arrow_forwardYou have two E. coli strains, XL10 and K12. XL10 is ampicillin resistant, or AmpR, and can grow on lab media plates in the presence of ampicillin. K12 is ampicillin sensitive, or AmpS, and cannot grow on lab media plates in the presence of ampicillin. You suspect that the AmpR gene is carried on a plasmid in XL10, rather than on its chromosome, and want to design an experiment to test this hypothesis. In designing your experiment, you may assume that you have access to the following materials: all reagents, plates, plasmids, and equipment used in the lab this semester Overnight cultures of XL10 & K12 Competent cells of XL10 & K12 a. Describe briefly the steps of a simple experiment that you could perform to test the hypothesis that the AmpR gene is carried on a plasmid in XL10 and not on its chromosome. (NOTE: no need for detailed procedure, simply state what you would do is sufficient). b. What specific experimental result will tell you that your hypothesis is correct?arrow_forwardYou transform bacteria with a plasmid carrying the ampicillin-resistance gene ampR. How would you determine which bacteria took up the plasmid? O Bacteria containing the plasmid would be able to grow in the absence of ampicillin. O Bacteria containing the plasmid would be able to grow in the presence of ampicillin. O Bacteria containing the plasmid would be able to produce ampicillin. O Ampicillin-resistance is necessary for the plasmid DNA get into the bacteria.arrow_forward
- Some DNA vaccines use a brief and small electrical shock to get DNA in plasmids into cells. What advantage would there be in using DNA vaccines that consist of plasmids instead of just pieces of double-stranded DNA? The new Covid19 vaccine produced by two companies (Pfizer, Moderna) uses mRNA coding for part of the spike protein of the virus. The virus uses the spike protein to invade human cells where it replicates. Is it surprising that the mRNA must be stabilized with chemicals that need ultra-cold or frozen storage to protect the mRNA from degradation before it causes human muscle cells to make the spike protein? Why not just inject the double-stranded cDNA that codes for the spike protein of the virus? What additional step or steps would you need to use to get the human muscle cells to produce the spike protein if the cDNA was injected to serve as the virus?arrow_forwardIn several experiments, you find these orders of genes getting transferred in from the chromosome with integrated F plasmid. Note the experimental results are listed vertically this time. Expt3 K Exptl Expt2 Expt4 Expt5 A J E D F N B M L H F P A I I H C J H L Use this data to create a map. . draw the correct "shape" of a map.arrow_forwardAntibiotic resistance genes are often found on R plasmids (see Natural Gene Transfer and Antibiotic Resistance). A likely source of the R plasmids is bacteria that produce the antibiotic. Why would some bacteria produce antibiotics (chemicals that kill bacteria) and why would they carry R plasmids?arrow_forward
- Agrobacterium tumefaciens is a bacterium that infects a wide range of broad-leaved plants. During infection, a plasmid called Ti is transferred into the plant cells, where that plasmid DNA is integrated into the plant’s genome. Some of the genes encoded by this plasmid are expressed by host cell machinery in the plant cell. These gene products produce plant hormones which stimulate cell division in the infected cells, causing the development of a plant tumor which provides an environment for that bacterium to grow. You are studying a novel isolate of A. tumefaciens and want to determine which of the genes on the Ti plasmid are expressed in the bacterium, and which are expressed in the plant. You start by sequencing the Ti plasmid. a) Considering the DNA sequence of Ti, describe two specific genetic features that would help you to differentiate whether a given gene is likely expressed in the plant or bacterium. b) You collect plant tumor tissue, containing both bacterial and plant…arrow_forwardA researcher has a Trp− auxotrophic strain of E. coliwith a mutation in a single gene. To identify thatmutant gene, she uses a genomic library made from a wild-type version of that same strain to find plasmids that rescue the mutant phenotype. The result issurprising. She recovers 10 plasmids that provide aTrp+ phenotype, but six of the plasmids contain geneX, while the other four contain gene Y. Our scientisthas encountered a phenomenon called multicopysuppression, related to the fact that plasmids are usually present in several copies per bacterium. Becausethe genes in the plasmids are present in more thantheir usual single copy in the bacterial chromosome,more than the usual amount of Protein X or Protein Yis being produced from the plasmids. Sometimes,overexpression of one protein can rescue the mutantphenotype caused by loss of a different protein.Suggest at least two ways that our scientist could determine which of the two genes, gene X or gene Y, actually corresponds to the…arrow_forwardWe transformed E coli cells with a plasmid modified to contain a ‘virulence factor’ which would allow growth on media containing the antibiotic kanamycin (Kan). The plasmid confers constitutive resistance to ampicillin (Amp). The bacterial experiment is about understanding whether such a ‘virulence factor’ confers physiological adaptation to Kan or whether the development of resistance can be explained by random mutations. For each independent transformation we re-suspended the cells from three colonies in Luria broth. For each suspension of cells we plated 100 microliters on a Kan plate. To estimate the number of cells seeded on each Kan plate we made four serial dilutions that were plated on Amp plates (1 – 4) and we counted the number of cells growing on them. From this we extrapolated how many cells had been seeded on the Kan plate. Then we normalised the Kan results for all the plates, assuming that every plate had been seeded with 10[5] cells. Consider two Kan plates, each with…arrow_forward
- Plasmids are important for recombining DNA. B.) What are the requirments of an expression vector.arrow_forwardA plasmid, pUC18, contains the ampicillin-resistance gene, the origin of replication, and the ß - gal gene, which codes for the B-galactosidase protein. This protein can break down the synthetic chemical X-gal, producing a blue product that stains the entire cell blue (but is harmless to the bacteria). At the beginning of the B-gal gene there are several unique restriction sites (some of them are shown in the diagram below). You wish to clone a 1.0-kb Xbal fragment into the pUC18 plasmid, so you cut the plasmid with Xbal and, after removing the enzyme, mix the Xbal-cut plasmid with the 1.0-kb fragment, ligate, and transform competent bacteria. Pati Xbal EcoRI B-gal A Amp ori Figure: pUC18 plasmid map (a) On what medium would you grow your transformed bacteria? (b) Do you expect the bacteria carrying plasmid pUC18 (without the insert) to be blue or white when grown in the presence of X-gal? Explain.arrow_forwardYou have set up a recombinant DNA experiment using the plasmid PBR322 as the vector (see plasmid below). You use the BamHI restriction site on the plasmid to insert the target DNA. The plasmid is then used to transform E.coli colls Is the following statement True or False? Growth of the transformed cells on agar containing both ampicillin and tetracycline will eliminate any cells that do not contain a plasmid. Clal Hindlll EcoRI Pvul BamHI Pstl amp tet PBR322 -Sall ori rop Pvull True Falsearrow_forward
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