Test Bank For Genetic Analysis An Integrated Approach by Sanders

Test Bank for Genetic Analysis An Integrated Approach by Sanders
Test Bank for Genetic Analysis An Integrated

Approach by Sanders
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Genetics: An Integrated Approach (Sanders)
Chapter 6 Genetic Analysis and Mapping in Bacteria and Bacteriophage
6.1 Multiple-Choice Questions
1) Why does conjugation with an Hfr donor result in a much higher rate of gene transfer than
conjugation with an F+ donor?
A) An F+ donor makes fewer pili.
B) An F+ donor's T strand is slow to reach the pilus.
C) The Hfr donor transfers genes from the chromosome and not the plasmid.
D) The Hfr donor segment must undergo recombination in the recipient.
E) The F+ donor segment must undergo recombination in the recipient.
Answer: C
Section: 6.1
Skill: Synthesis/Evaluation
2) In phi X 174 phage, whose genome is a single-stranded, circular, DNA known as the - strand,
replication occurs by the rolling circle method. In which order must the events of replication occur?
A) nicking of the - strand, binding of relaxase protein to the 5′ end, DNA synthesis of a + strand,
separation of + from - strands, circularization of both strands
B) synthesis of a + strand, nicking at the origin, binding of protein to the 5′ end, DNA synthesis,
displacement of the + strand when replication passes the origin, circularization of the + strand
C) synthesis of a + strand, nicking of both strands to separate them, recircularization of both strands
D) binding of a protein at the origin of the - strand, synthesis of a + strand, release of the + strand
Answer: B
Section: 6.1
3) A large region of the F plasmid (~33 kb) is called the transfer region (tra) and contains about 40
genes necessary for conjugation. The gene traA codes for the protein pilin that is polymerized to form a
pilus, using a number of other tra genes. The pilus is a hollow cylinder whose units disassemble once
contact is made in order to "reel in" the recipient cell. What other functions of the tra genes are required
for transfer?
A) DNA unwinding, and restriction of contact with F+ cells
B) DNA replication enzymes
C) resistance to phage infection
D) integration of F genes into the recipient's chromosome
E) integration of F genes into the recipient's plasmid
Answer: A
Section: 6.1
Skill: Application/Analysis
1
Copyright © 2013 Pearson Education, Inc.
4) When F+ conjugation occurs successfully, only one copy of the F plasmid single-stranded DNA is
transferred. What else must minimally occur?
A) circularization of the recipient's new DNA
B) circularization plus replication of the new recipient DNA
C) stopping the rolling circle replication after one copy length, circularization, and replication
D) continuation of rolling circle replication in the donor cell and replication in the recipient cell
E) rolling circle replication in the recipient cell
Answer: C
Section: 6.1
5) To integrate an F plasmid's genes into a host chromosome, there must be DNA recombination at
homologous regions on the two circular strands. What is the minimum number of such crossover events
needed for integration?
A) 1
B) 2
C) 3
D) 4
E) 8
Answer: A
Section: 6.1
6) When F+ conjugation occurs successfully, only one copy of the F plasmid single-stranded DNA is
transferred. What else must minimally occur?
A) circularization of the recipient's new DNA
B) circularization plus replication of the new recipient DNA
C) stopping the rolling circle replication after one copy length, circularization, and replication
D) continuation of rolling circle replication in the donor cell and replication in the recipient cell
E) rolling circle replication in the recipient cell
Answer: C
Section: 6.1
Refer to this text to answer the following questions.
To map genes of a bacterial strain, conjugation must be interrupted at given times. Suppose you have
Hfr cells of genotype a+b+c+d+e+strR and F= cells of genotype a-b-c-d-e-strS and you combine these
two cultures in liquid medium in four blenders at time 0. After intervals of 3, 6, 9, and 12 minutes, you
turn on successive blenders.
7) What role does the blender play in the experiment?

A) serves as a culture container only
B) acts as a mechanism to bring cells into contact
C) speeds up molecular movement of DNA
D) shears the sex pili of conjugants
E) hastens conjugation events
Answer: C
Section: 6.2
2
8) The resulting cultures were then plated on medium containing streptomycin. Why?
A) to eliminate non-conjugating Hfr cells
B) to eliminate all non-conjugated cells
C) to eliminate non-conjugated F- cells
D) to selectively eliminate cells that have taken in F+ genes
Answer: C
Section: 6.2
9) This graph shows the results of plating the cultures of each blender. What is the correct order of the
genes on this chromosome?
A) a b c d e
B) a c e b d
C) b e c a (d is unknown)
D) d a c e b
E) b e c a d
Answer: B
Section: 6.2
10) Which of the following allows the compilation of time-of-entry maps?

A) the use of many replicates of each experiment
B) the use of appropriate controls repeatedly
C) the fact that gene entry can proceed in either direction
D) the observation that Hfr genes recombine so efficiently
E) the recognition of phenotype interaction
Answer: C
Section: 6.3
3
11) In five Hfr strains, each of which was used to build a time-of-entry map, the genes entered the
recipient cells as follows:
Strain 1: S L A C T F
Strain 2: N P F T C A
Strain 3: T F P N U Y
Strain 4: S H Y U N P
Strain 5: U N P F T C
Which of the following represents a correct compilation of these results?
A) S L A C T F P N H C U
B) N P F T S L A C H U T
C) T C A L S P N U Y H
D) U N P F T C A L S T F
E) S L A C T F P N U Y H
Answer: E
Section: 6.2
12) An F′ donor includes F DNA plus a segment of bacterial chromosome DNA. If conjugation is
interrupted before the entire F′ chromosome transfers, what could be the expected consequences?
A) integration of some but not all F′ genes into the recipient's chromosome
B) a smaller F′ plasmid than had been present in the donor
C) failure of the F′ strand to circularize in the recipient
D) change of the recipient from F- to F+
E) formation of a recipient that is a stable partial diploid
Answer: C
Section: 6.3
13) In a partially diploid F′ exconjugant, there are two alleles for lac (1 and 2) and two alleles for his (3
and 4). The phenotype of this cell is identical to a lac 1 his 4 cell. What is the best interpretation?
A) Lac 1 and 2 and his 3 and 4 share only lac 1 and his 4 phenotypes.
B) Lac 1 complements lac 2 and his 3 complements his 4.
C) Lac 1 and his 4 are dominant.
D) Lac 1 and his 4 are wild types.
E) Lac 1 shows its phenotype only if his 4 is also present.
Answer: C
Section: 6.3
4
14) A competent Hemophilus strain is individually transformed by three genes (a, b, and c) at a rate of
0.002% each. However, cotransformation frequencies for each pair are as follows:
a and b 0.38 %
a and c 0.72%
b and c 0.19%
What is the correct conclusion from this data?
A) Genes b and c are farthest apart.
B) Gene order is c, b, a.
C) Cotransformation of b and c is so frequent, they must be one gene.
D) Gene a is closer to b than to c.
E) Genes a and c are farthest apart.
Answer: A
Section: 6.4
15) In his analysis of rII in phage, Benzer assayed complementation. Suppose you try your hand at this
with several mutants and get the following results (+ = complementation):
42 and 62 +
42 and 83 -
62 and 63 -
62 and 74 +
42 and 74 -
42 and 75 +
62 and 75 -
Mutation 42 has previously been shown to be in a different gene than mutation 62. What can you
conclude?
A) Mutations 42 and 75 are in the same gene.
B) Mutations 62 and 74 are in the same gene.
C) Mutations 62, 63, and 74 are in the same gene.
D) Mutations 42 and 74 are in the same gene.
E) Mutation 63 is in a gene with neither 62 nor 42.
Answer: D
Section: 6.6
16) In generalized transduction, a phage introduces a segment of donor DNA into the recipient cell. This
is followed by recombination of the donor fragment with the recipient chromosome. Which of the
following must occur?
A) circularization of the donor fragment before recombination
B) a pair (or even number) of crossovers between the donor segment and the recipient
C) degradation of one of the two strands of phage genome
D) replication of the donor segment before recombination
E) a single crossover between the donor segment and the recipient chromosome
Answer: B
Section: 6.5
5
17) In an imagined transduction experiment with alleles a+, b+, and c+, a+ alone was transduced into
an auxotroph 1400 times, b+ alone 1200 times, and c+ alone 970 times. Genes a+ and b+ were both
found 10 times, b+ and c+ 2 times, and a+ and c+ one time. What can you conclude?
A) Cotransduction is too infrequent to be used to map genes.
B) Genes a and c cotransduce only accidentally.
C) Genes a and b are more closely linked than a and c or b and c.
D) Genes a and c are found together only if double crossovers have occurred.
E) Transduction of gene c is very rare.
Answer: C
Section: 6.5
Refer to this text to answer the following questions.
Bacterial gene transfer is also used in the lab to introduce genes into organisms of different species,
genera, phyla, or even kingdoms or domains. A major example is the use of a Ti plasmid of the soil
bacterium Agrobacterium tumefaciens to transfer genes into dicotyledonous plants. The bacterium
infects the plant root in the soil and transfers some of the Ti plasmid genes (T DNA) into the plant
nucleus. Recombination may occur, and T DNA expression is controlled by plant compounds generated
in response to wounding and infection.
18) The T DNA is generated when there is a nick that creates a primer for replication, followed by
transfer of a single-stranded DNA piece that converts to a double-stranded piece in the plant nucleus.
Which of these processes is Ti plasmid transfer most like?
A) conjugation
B) generalized transduction
C) specialized transduction
D) transformation
Answer: A
Section: 6.1
19) This method can be used to introduce a gene for an antigenic protein into an edible plant, such as a
potato, to produce an edible vaccine (e.g., for cholera). What has to be made a part of the Ti plasmid?
A) a gene for cholera resistance
B) a gene for a subunit of cholera toxin, including its regulatory elements
C) a gene for that part of the human immune system that neutralizes cholera
D) a molecule of anti-cholera antibody
E) a polypeptide that can attach the T DNA to plant DNA
Answer: B
Section: 6.3
6
20) Assuming you have appropriately engineered the Ti plasmid to carry the gene in question, how
could you best and most safely test whether your potatoes are now working as a vaccine?
A) Feed the engineered potato to a population at high risk for cholera.
B) Cook the potatoes to denature the toxin and feed them to mice susceptible to cholera.
C) Feed pieces of raw potato to susceptible mice and test them for antibodies.
D) Emulsify the potatoes in a liquid that can be injected into experimental animals.
Answer: C
Section: 6.3
21) Benzer's analysis of phage genomes included deletion mapping. Since recombination could not
occur in the area of a deletion, an infection by two phage strains, one of which has a deletion and the
other a point mutation, which results in no wild-type recombinants being produced indicates ________?
A) that the deletion includes the position of the point mutation in the same gene
B) that the deletion includes the position of the point mutation in a different gene
C) that the point mutation is dominant
D) that the point mutation is recessive
E) the distance between the two types of mutation
Answer: A
Section: 6.6
6.2 Short-Answer Questions
1) Name the two cycles in which bacteriophages act when they infect bacteria.
Answer: lysis and lysogeny
Section: 6.5
Skill: Knowledge/Comprehension
2) What is the difference between F+ and F- bacteria?

Answer: F+ bacteria have an F plasmid.
Section: 6.1
3) Describe two features of a plasmid.

Answer: two of the following: circular, double-stranded DNA, autonomous, or includes genes
Section: 6.1
4) Which form of bacterial recombination requires no vector?

Answer: transformation
Section: 6.4
5) Name the three usual phases of bacterial growth in a liquid medium.

Answer: lag (phase), log (phase), stationary (phase)
Section: 6.1
7
6) If you count 73 plaques on a bacterial plate when you had added 0.1 ml of a 10-4 dilution of phage,
what was the initial concentration of the (undiluted) phage?
Answer: 73 × 104 × 10 = 7.3 × 106 pfu/ml
Section: 6.5
7) In a bacterial cross of an Hfr donor a+b+ with a recipient F- a-b-, what would be a possible
recombinant?
Answer: a+b- or a-b+
Section: 6.2
8) In a transduction experiment, what would constitute evidence of linkage?

Answer: cotransduction (of genes)
Section: 6.5
9) All three types of bacterial recombination share one characteristic. What is it?
Answer: one-way transfer of gene(s), or transfer from donor to recipient
Section: 6.1
10) One kind of plasmid in bacteria is called an R plasmid. What kinds of genes are typical of such
plasmids?
Answer: antibiotic-resistant genes
Section: 6.1
11) What is the role of the relaxosome in conjugation?

Answer: binds to F origin, or cleaves T strand, or binds to 5′ end to begin transfer
Section: 6.1
12) In conjugation, as the 5′ end of the T strand begins to move across the pilus, what form of replication
occurs in the donor cell?
Answer: rolling circle (replication)
Section: 6.1
13) In Hfr transfer, what happens to linear DNA that does not recombine with the host chromosome?
Answer: (enzymatic) degradation
Section: 6.1
14) If conjugation occurs with an F′ donor, how will the recipient be characterized genetically?
Answer: partially diploid
Section: 6.3
8
15) What characterizes a bacterial cell that can undergo transformation?
Answer: competence, or ability to take up double-stranded DNA
Section: 6.4
16) What distinguishes generalized versus specialized transduction?

Answer: generalized uses lytic phage; specialized uses lysogenic phage
Section: 6.5
17) Jacob and colleagues developed a bacterial linkage map based on time. What kind of recombination
was used?
Answer: conjugation
Section: 6.2
18) In an E. coli conjugation experiment, the donor cell is found to be thr+, leu+, his+, and the F- cell is
thr-, leu-, and his-. When the mating is interrupted after 10 minutes and the recipients are plated, they
are found to be leu+ and his+ but thr-. What can you conclude?
Answer: Leu and his are close together.
Section: 6.2
19) In a conjugation experiment, the Hfr donor is thr+, leu+, his+, str. The recipient is thr-, leu-, and
his- strS. Why would you use medium containing streptomycin to analyze your results?
Answer: to identify strR recipients
Section: 6.2
20) Arabinose sugar is used by ara+ bacteria but cannot be used by ara- mutants. When both ara3 and
ara5 mutants are introduced into the same recipient via a vector, and the recipients still cannot use
arabinose, what can you conclude?
Answer: The mutations are in separate genes.
Section: 6.6
21) Seymour Benzer's fine structure studies of genomes used mutations found in specific regions of
what kind of genome?
Answer: bacteriophage (phage)
Section: 6.6
22) What is meant by a mutation that Benzer referred to as nonrevertible?

Answer: due to a deletion
Section: 6.6
9
6.3 Fill-in-the-Blank Questions
1) Some bacteriophages can be either lytic or lysogenic. These are called ________ phages.
Answer: temperate
Section: 6.5
2) Prototrophs are bacteria that can grow on ________ medium.

Answer: minimal
Section: 6.1
3) Once a phage genome has become part of the host chromosome, it is called a ________.
Answer: prophage
Section: 6.5
6.4 Essay Questions
1) When a soil bacterium infects a plant root and introduces its Ti plasmid, how does this affect both the
infectious bacterium and the plant?
Answer: A soil bacterium, Agrobacterium tumefaciens, contains a tumor-inducing (Ti) plasmid that is
~200 kb and contains the T-DNA and the genes required to transfer that DNA to the plant cell.
Bacterial plasmids are circular, so DNA must be linearized in order to be able to incorporate into the
plant's linear chromosomes. When A. tumefaciens infects the plant, the bacterium must cut the T-DNA
out of the circular plasmid. DNA is nicked at the right border sequence, yielding a region with single-
stranded DNA. Single-stranded binding proteins attach to this single stranded DNA. DNA synthesis will
displace the single-stranded region and a second nick at the left border region will release the single
stranded T-DNA fragment. Through the process of single strand DNA transfer (similar to bacterial
conjugation), this fragment can be transferred to the plant cell. The DNA is targeted to the plant cell
nucleus, where it can be incorporated into the host genome. Through homologous recombination, the
DNA becomes integrated into the host genome, where the gene products will be transcribed and
translated as any other plant gene by the plant's own transcription and translation machinery.
Section: 6.1
10
2) Conjugation, transduction, and transformation all occur in nature. Which do you expect would more
likely account for the acquisition of the genes for resistance to an antibiotic? Why? Which do you think
would best account for the acquisition of genes from one bacterial species by another of a different
genus? Why?
Answer: Through the process of lateral gene transfer, genes located on a circular strand of DNA called
an R-plasmid contain antibiotic-resistant genes. Through conjugation, an antibiotic-resistant bacterium
can transfer the R-plasmid to a non-resistant bacterium. Because this R-plasmid provides a selective
growth advantage to some bacteria, they may "collect" many copies of various antibiotic resistance
genes from other bacteria.
Transfer of genes between bacterial species likely involves transformation, the process by which
extracellular fragments of DNA released when a donor bacterial cell lyses can be absorbed across the
cell membrane of a competent recipient cell as transforming DNA.
Specialized transduction involves bacteriophages, which can transfer host DNA from one host to the
next through aberrant excision of a lysogenic prophage. However, it is more likely that gene transfer
between species occurred through transformation rather than transduction.
Section: 6.5
3) Which, if any, characteristics of bacterial gene transfer is/are useful in analyzing eukaryotic gene
transfer?
Answer: Lateral gene transfer is the process by which genes are transferred between species, as opposed
to vertical gene transfer, in which genes are inherited from ancestors. This phenomenon is common in
prokaryotes, and can occur by conjugation, transformation, and transduction. There is evidence of
natural lateral gene transfer in eukaryotes, but it is far more relevant in terms of artificial gene transfer
and genetic engineering. Genes from one organism can be successfully expressed in a different
organism, e.g. expression of GFP, luciferase, and various reporter genes. If you want to introduce a gene
of interest into a eukaryotic cell, you will most often use transformation. Cells are incubated with the
DNA of interest, and then subjected to artificial selection to select for cells that have taken up the DNA
from the environment. This same process occurs naturally in bacteria and in single-celled eukaryotes,
but can be a valuable asset for genetic engineering.
More recently, specially engineered viruses have also been used to introduce DNA into a eukaryotic cell
as a strategy for creating transgenic cell lines or for gene therapy. This process is similar to the
mechanism by which bacteriophages transfer DNA between bacteria through the process of
transduction.
Thus, studying the mechanisms of lateral gene transfer is both helpful and relevant to understanding the
processes by which eukaryotes acquire new genes naturally or through intentional genetic engineering.
Section: 6.1
11
Test Bank for Genetic Analysis An Integrated Approach by Sanders
4) How are selective media used in studying bacterial recombination?

Answer: Selective media permits certain bacteria to grow while suppressing the growth of other
bacteria. During bacterial recombination, you can accurately assess transfer of genes using selective
media in two ways.
First, you can use selective media to identify bacteria that have acquired a resistance gene and are
recombinant. For example, E. coli can be transformed with a plasmid containing your gene of interest
and a selectable marker, often an antibiotic resistance gene. You can add antibiotic to a bacterial culture
and only the bacteria that have taken up the plasmid will be resistant. This will allow for identification
of sequences which have been transferred between bacteria.
Second, you can use selective media to identify bacteria which have a mutation in a metabolic pathway
by looking for mutants that are unable to complete normal metabolic processes. For instance, let's say
you are studying a metabolic pathway and wish to identify bacteria that have mutations in this pathway.
You can grow the bacteria in media with and without the final product of the pathway. If the pathway is
mutated and one of the enzymes is absent, the bacteria must be supplemented with certain nutrients to be
able to grow. This will allow you to identify transformants, or mutants of interest.
Section: 6.1
5) Explain when, why, and how recombination in bacteria is like crossing over in diploid cells.
Answer: Both bacterial recombination and crossing over during meiosis require several factors. First,
both processes require nicking of DNA. In bacteria, this is when the DNA is nicked in preparation for
the transfer of single-stranded DNA via conjugation. In eukaryotes, one strand of the DNA is nicked
during the formation of chiasmata. This allows for strand invasion and crossing over between
homologous chromosomes.
Second, both processes use similar enzymes to mediate recombination. DNA helicases are involved in
relieving torsional strain in the circular and linear chromosomes. Ligases are necessary for repairing the
phosphodiester bonds within the DNA backbone that were created by the single-stranded breaks. Single-
stranded binding proteins (SSBs) assist in stabilizing the otherwise unstable ssDNA and allow it to
undergo processes such as replication, recombination, and transcription.
Third, both processes require pairing of homologous sequences. In bacteria, when the chromosome
DNA from the donor cell is transferred to a recipient bacterium, the homologous parts of the donor and
recipient DNA molecules can undergo recombination that leads to a change in the genotype of the
recipient cell. In eukaryotes, proper pairing of homologous sequences is necessary for even exchange of
chromosomes during crossing over. If homologous sequences are unevenly pairing, this can result in a
region of chromosome being deleted or duplicated in the resulting gametes.
Section: 6.4
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Test Bank for Genetic Analysis An Integrated

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6.1 Multiple-Choice Questions

Refer to this text to answer the following questions.

7) What role does the blender play in the experiment?

10) Which of the following allows the compilation of time-of-entry maps?

Refer to this text to answer the following questions.

6.2 Short-Answer Questions

2) What is the difference between F+ and F- bacteria?

3) Describe two features of a plasmid.

4) Which form of bacterial recombination requires no vector?

5) Name the three usual phases of bacterial growth in a liquid medium.

8) In a transduction experiment, what would constitute evidence of linkage?

11) What is the role of the relaxosome in conjugation?

16) What distinguishes generalized versus specialized transduction?

22) What is meant by a mutation that Benzer referred to as nonrevertible?

2) Prototrophs are bacteria that can grow on ________ medium.

6.4 Essay Questions

4) How are selective media used in studying bacterial recombination?

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