ND STONY BROOK UNIVERSITY LOOSELEAF GENETICS: FROM GENES TO GENOMES
6th Edition
ISBN: 9781260406092
Author: HARTWELL, Leland, HOOD, Leroy, Goldberg, Michael
Publisher: Mcgraw-hill Education/stony Brook University
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Chapter 14, Problem 24P
You can carry out matings between an Hfr and F strain by mixing the two cell types in a small patch on a plate and then replica plating to selective medium. This methodology was used to screen hundreds of different cells for a recombination-deficient recA- mutant. Why is this an assay for RecA function? Would you be screening for a recA mutation in the F or Hfr strain using this protocol? Explain.
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After transformation you were asked to grow bacterial cells transformed with plasmid on a plate that had X-gal and ampicillin. X-Gal is often used as in indicator dye, which turns blue when metabolized by B-galactosidase protein and used to test if cloning experiments have worked. [Note look at the vector diagrams carefully]
Briefly explain how you would find the bacterial cells that are transformed with the plasmid with the YFG inserted.
A prototrophic strain (his" arg" lac') was used as a donor to transform an auxotrophic strain (his arg
lac'). Initial transformants are isolated on minimal medium + histidine + arginine + lactose - glucose.
i.
What genotypes will grow on this medium?
ii.
These colonies are replicated to minimal medium + histidine, and 50% of the original
colonies grow. What genotypes will grow on this medium?
iii.
The original colonies are also replicated to minimal medium + arginine, and 10% of the
colonies grow. What genotypes will grow on this medium?
iv.
The original colonies are also replicated to minimal medium. No colonies grow. Based
on this information, what genotypes will grow on minimal medium + histidine and on
minimal medium + arginine? What is the relative gene order for his, arg, and lac? Which
two genes are closer? Explain your answer.
Assume you have successfully cloned a small (200 bp) fragment of DNA into the polylinker region of a pUC18 cloning vector. Describe the appearance of transformed colonies you would expect to see on each of the following plates: plain media, media containing ampicillin, media containing tetracycline, media containing ampicillin and X-Gal.
Chapter 14 Solutions
ND STONY BROOK UNIVERSITY LOOSELEAF 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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- 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…arrow_forwardA plaque assay is performed beginning with 1 mL of a solution containing bacteriophages. This solution is serially diluted three times by combining 0.1 mL of each sequential dilution with 9.9 mL of liquid medium. Then 0.1 mL of the final dilution is plated in the plaque assay and yields 17 plaques. What is the initial density of bacteriophages in the original 1 mL?arrow_forwardIn a generalized-transduction experiment, phages arecollected from an E. coli donor strain of genotype cys+leu+ thr+ and used to transduce a recipient of genotypecys- leu- thr-. Initially, the treated recipient populationis plated on a minimal medium supplemented with leucine and threonine. Many colonies are obtained.a. What are the possible genotypes of these colonies?b. These colonies are then replica plated onto threedifferent media: (1) minimal plus threonine only, (2)minimal plus leucine only, and (3) minimal. Whatgenotypes could, in theory, grow on these three media?c. Of the original colonies, 56 percent are observed togrow on medium 1, 5 percent on medium 2, and nocolonies on medium 3. What are the actual genotypes ofthe colonies on media 1, 2, and 3?d. Draw a map showing the order of the three genes andwhich of the two outer genes is closer to the middle genearrow_forward
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