chapter 3

Introduction to Biotechnology, 4e, Global Edition (Thieman)
Chapter 3 Recombinant DNA Technology and Genomics
1) You just cloned a new gene from mice. Which of the following techniques would be the best
choice to use if you wanted to determine the number of copies of this gene in the mouse
genome?
A) Northern blot analysis
B) RT-PCR
C) Western blot analysis
D) Southern blot analysis
E) In situ hybridization
Answer: D
Section: 3.3 Laboratory Techniques and Applications of Recombinant DNA Technology
Bloom's Taxonomy: Remembering/Understanding
2) Which of the following techniques involves hybridizing a cDNA sample to a chip containing
thousands of single-stranded DNA sequences, allowing one to study the expression of thousands
of genes simultaneously?
A) PCR
B) Southern blot
C) FISH
D) Agarose gel electrophoresis
E) DNA microarray
Answer: E
3) When making a complementary DNA (cDNA) library, which enzyme is used to copy mRNA
into DNA?
A) DNA ligase
B) DNA polymerase
C) Primase
D) RNA polymerase
E) Reverse transcriptase
Answer: E
Section: 3.2 How Do You Identify and Clone a Gene of Interest?
1
Copyright © 2020 Pearson Education Ltd.
4) In a recombinant DNA experiment, which enzyme is used to join together DNA fragments by
forming phosphodiester bonds between nucleotides? Recall that this same enzyme joins together
Okazaki fragments on the lagging strand during DNA replication.
A) DNA primase
B) DNA polymerase
C) DNA helicase
D) DNA ligase
E) Reverse transcriptase
Answer: D
5) During library screening, PCR, Southern blotting, and other techniques, binding two pieces of
DNA to each other by hydrogen bonding is called ________.
A) autoradiography
B) hybridization
C) DNA ligation
D) reverse transcription
E) polyadenylation
Answer: B
6) ________ is a DNA-binding dye that fluoresces when DNA in an agarose gel is illuminated
with ultraviolet light.
A) Casein
B) Ethidium bromide
C) Malt
D) Lactic acid
E) Ethanol
Answer: B
7) Which of the following vectors would be the best choice for gene transfer in plant cells?
A) Cosmid
B) Bacterial artificial chromosome
C) Bacteriophage vector
D) Ti vector
E) Plasmid
Answer: D
Section: 3.1 Introduction to Recombinant DNA Technology and DNA Cloning
2
8) A ________ is a single-stranded DNA molecule attached to a radioactive or fluorescent
compound that is complementary to a specific sequence of DNA. Such pieces of DNA are used
to identify and study cloned genes in hybridization experiments.
A) polypeptide
B) promoter
C) vector
D) probe
E) plasmid
Answer: D
9) Transformation in a cloning experiment is ________.

A) ligating pieces of foreign DNA together
B) inserting DNA into bacteria cells
C) cutting DNA with restriction enzymes
D) using PCR to clone a gene
E) a technique for determining gene copy number in a genome
Answer: B
10) Which of the following is an INCORRECT statement about restriction enzymes?

A) Most restriction enzymes are isolated from bacteria.
B) Restriction enzymes usually recognize palindromic sequences.
C) Restriction enzymes create phosphodiester bonds between pieces of DNA in a cloning
experiment.
D) Restriction enzymes can cut to create overlapping single-stranded ends of DNA.
E) Restriction enzymes can cut to create blunt-ended pieces of DNA.
Answer: C
11) Based on what you know about which organisms naturally produce restriction enzymes and
how restriction enzymes are named, which of the following enzymes is isolated from the
bacterium Bacillus amyloliquefaciens?
A) EcoRI
B) BamHI
C) SmaI
D) PstI
E) HindIII
Answer: B
Bloom's Taxonomy: Applying/Analyzing
3
12) Imagine you wanted to use a human genomic DNA library to clone the human gene for
insulin. You will be using the rat insulin gene sequence as your DNA probe. In what order would
you perform the following steps to accomplish this goal?
1. Use autoradiography to identify colonies containing DNA that hybridized to the probe
2. Grow transformed cells on media with antibiotics and X-gal for blue-white screening
3. Ligate genomic DNA and vector DNA
4. Cut genomic DNA and vector DNA with restriction enzymes
5. Hybridize library DNA with a labeled probe for the rat insulin gene
6. Transform bacteria with recombinant plasmid
A) 2, 3, 6, 5, 4, 1
B) 1, 2, 5, 6, 4, 3
C) 4, 3, 6, 2, 5, 1
D) 4, 3, 5, 2, 6, 1
E) 2, 6, 5, 3, 1, 4
Answer: C
13) Approximately how large is the human genome?

A) 20,000 bp
B) 1 billion bp
C) 3 billion bp
D) 13 million bp
E) 30 billion bp
Answer: C
Section: 3.4 Genomics and Bioinformatics: The Hottest Disciplines in the History of
Biotechnology
14) Approximately how many genes are present in the human genome?
A) 3 billion genes
B) 20 billion genes
C) 100,000 genes
D) 20,000 genes
E) 50,000 genes
Answer: D
Biotechnology
4
15) Which of the following techniques is the best choice for amplifying DNA?
A) Southern blot analysis
B) PCR
C) DNA sequencing
D) Affinity chromatography
E) Microarray analysis
Answer: B
16) Which of the following techniques is most commonly used to separate and analyze DNA by
size?
A) PCR
B) Agarose gel electrophoresis
C) DNA microarray analysis
D) Hybridization
E) DNA libraries
Answer: B
17) A ________ consists of cloned DNA fragments for all expressed genes in a particular tissue
A) genomic DNA library
B) cDNA library
C) PCR library
D) Guggenheim library
E) shotgun library
Answer: B
18) Which of the following techniques is most efficient for transforming bacterial cells?
A) Gene gun
B) ES cell transfer
C) Electroporation
D) Pronuclear microinjection
E) Sperm-mediated transfer
Answer: C
5
19) Which of the following statements about the human genome is INCORRECT?
A) There are approximately 3.1 billion bp in the human genome.
B) Humans share a majority of genes with other species.
C) Less than 2 percent of the genome codes for proteins.
D) The human genome contains approximately 100,000 genes.
E) The functions of most human genes are still unknown.
Answer: D
Biotechnology
20) Dideoxyribonucleotides (ddNTPs) used for DNA sequencing lack oxygen atoms at
________.
A) the 1' carbon of the pentose sugar
B) the 1' and 2' carbons of the pentose sugar
C) the 2' and 3' carbons of the pentose sugar
D) the 3' and 5' carbons of the pentose sugar
E) the 5' carbon of the pentose sugar
Answer: C
21) A recombinant DNA molecule produced artificially contains sequences ________.

A) from bacteria only
B) from yeast only
C) from unrelated organisms
D) from cells that have been chemically modified
E) from prokaryotic genes
Answer: C
Section: 3.5 Systems Biology and Synthetic Biology
22) A linear strand of DNA is 1,000 bp long. A recognition sequence for the restriction enzyme
Eco R1 is located 300 base pairs (bp) from the 5' end of this linear DNA molecule. Digesting this
DNA molecule with Eco RI would produce ________.
A) one DNA fragment, 1,000 bp long
B) three DNA fragments, two of them 300 bp long and one 400 bp long
C) two DNA fragments, one 300 bp long and one 700 bp long
D) two fragments, each 500 bp long
E) none of the above
Answer: C
6
23) During molecular cloning, a gene of interest (insulin) is inserted into a bacterial structure
called a ________ and enters a bacterial cell through a process called ________.
A) chromosome; electrophoresis
B) nucleus; transformation
C) plasmid; transcription
D) plasmid; transformation
E) genome; transformation
Answer: D
24) Which of the following might lead to the generation of several incorrect PCR products?
A) An annealing temperature that is too low
B) An annealing temperature that is too high
C) Performing 30 cycles of PCR
D) Using the correct primers
E) Including an extension step
Answer: A
25) A bacteriophage is ________.

A) a small cloning vector used in humans
B) a large cloning vector used in humans
C) a type of bacterial protein
D) a virus that specifically infects bother viruses
E) a virus that specifically infects bacteria
Answer: E
26) Explain the use of an antibiotic (e.g., ampicillin) resistance gene on a vector.
Answer: The antibiotic resistance gene is found on the vector (also known as the plasmid). This
gene confers resistance to the recombinant DNA plasmid when transformed into bacterial cells
and plated on agar media containing the antibiotic, such as ampicillin. Only bacterial cells that
have taken up the vector can grow on media containing ampicillin, thus allowing for a selection
of colonies that have taken up the vector.
27) Describe two advantages for cloning a gene from a cDNA library versus a genomic DNA
library.
Answer: A cDNA library is free of introns and it is enriched for the gene of interest because it is
made from mRNA from the desired tissue type. In contrast, a genomic DNA library still has
introns and it has every gene in the genome, making it more difficult to find the gene of interest.
7
28) Describe three reagents used and the three steps in the process of polymerase chain reaction.
Answer: The reaction calls for Taq polymerase, a pool of nucleotides, primers, and a thermo
cycler. The three steps in the process are as follows: denaturation, which occurs at 94°C to
separate the DNA strand; annealing, in which the temperature varies depending on the primers
but is usually between 50°C and 60°C and allows the primers to adhere to the DNA strands; and
extension, which occurs at 72°C, allowing for elongation of the DNA from the primer.
29) Explain the meaning and utility of RNA interference.

Answer: RNA interference (RNAi) is a process within living cells that moderates the activity of
their genes.
30) Explain the dideoxy sequencing method.

Answer: Sanger's method, which is also referred to as chain termination, is based on the use of
dideoxynucleotides (ddNTP's) in addition to the normal nucleotides (NTP's) found in DNA.
Dideoxynucleotides are essentially the same as nucleotides except they contain a hydrogen group
on the 3' carbon instead of a hydroxyl group (OH). These modified nucleotides, when integrated
into a sequence, prevent the addition of further nucleotides. This occurs because a
phosphodiester bond cannot form between the dideoxynucleotide and the next incoming
nucleotide, and thus the DNA chain is terminated. A nested set of products can then be used to
determine the actual nucleotide sequence.
31) Which of the following is NOT a feature of a DNA cloning Vector?

A) Origin of replication
B) DNA sequencing primer sequences
C) Selectable marker genes
D) DNA polymerase
E) Multiple cloning site
Answer: D
32) Which of the following methods is used to clone smaller pieces of a gene?
A) Southern blot
B) Western blot
C) Whole genome sequencing
D) PCR
E) 2D gel electrophoresis
Answer: D
8
33) Which of the following technologies is an example of third generation sequencing?
A) Single-molecule sequencing in real time
B) Whole genome shotgun sequencing
C) Real time PCR
D) High-throughput DNA sequencing
E) Sanger sequencing
Answer: A
34) The ________ is one technique that allows for the genome to be targeted at a specific
location and also allows for the removal of genes or insertion of genes in cultured cells or a
whole organism.
A) site-directed mutagenesis system
B) RNAi system
C) gene knockout system
D) CRISPR-Cas system
E) siRNA system
Answer: D
35) Which sequencing method allows for in situ analysis of gene expression and allow for
quantitative analysis of all RNAs in a cell?
A) High throughput sequencing
B) RNA-seq
C) Shotgun cloning
D) Whole-exome sequencing
E) Microarray
Answer: B
9
36) Describe a current technology used to sequence a single molecule of DNA. What are some
advantages and disadvantages of the technology?
Answer: The single-molecule sequencing in real time (SMRT) method is one technology used to
sequence a single molecule of DNA. SMRT works by attaching single-stranded molecules of the
DNA to be sequenced to a single molecule of DNA polymerase as it synthesizes a strand of
DNA. The DNA polymerase adds fluorescently tagged nucleotides to synthesize the DNA, and
the fluorescent tag is cleaved off each base as it is added to the DNA strand. The DNA
polymerase is confined within a hole that is in a thin layer of metal on a glass substrate, and such
a configuration allows for the detection of individual nucleotides as they are added to single
strands synthesized in the nanopore. Advantages include the ability to sequence and analyze
genomes from a single molecule in a short amount of time. Disadvantages include a somewhat
high error rate and they are expensive.
37) Describe a potential application for CRISPR-CAS and include in your description the basic
mechanism of CRISPR-CAS.
Answer: One potential application for CRISPR-CAS includes replacing an existing sequence of
DNA present in cells growing in culture with a new sequence of DNA, which would allow one to
study a phenotype or signaling a pathway of interest. To do this, the new sequence of DNA
would have a sequence that matches the existing sequence of DNA that needs to be edited, but
does not match any other sequence in the genome. Then the CRISPR associated nuclease Cas9
will cut the DNA at the site of interest.
38) How can the ENCODE project enhance our understanding of the genome?
Answer: The Encode findings have broadly defined the functional elements of the genome that
regulate the expression of human genes. This information has informed our understanding of the
genome because it has enhanced our knowledge of all the different functional elements (i.e.,
promoters, enhancers, transcriptional start sites) and their relationships, which allows for the
design of better drugs to treat diseases.
Biotechnology
39) ________ is a sequencing method that only sequences exons.

A) Whole-exome sequencing
B) Single-cell sequencing
D) High throughput sequencing
E) Exon-directed sequencing
Answer: A
Biotechnology
10
40) ________ is a sequencing method that involves isolating genomic DNA from a single cell to
analyze somatic mutations.
A) Whole-exome sequencing
B) Single-site sequencing
D) High throughput sequencing
E) Single-cell sequencing
Answer: E
Biotechnology
41) What is a network map? Describe how network maps can be used to help scientists model
potential interactions of molecules involved in normal and disease processes.
Answer: A network map is used in systems biology to show interacting proteins, genes, and
other molecules. They can be used to help scientists model potential interactions of molecules
involved in normal and disease processes because network maps generate computational models
that allow the running of simulations to determine how signaling events occur. This information
can inform scientists about potential drug targets and proteins that could be used for therapeutic
purposes.
Section: 3.5 Systems Biology and Synthetic Biology
11

chapter 3

Uploaded by

Copyright:

Available Formats

chapter 3

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

chapter 3

Uploaded by

Copyright:

Available Formats

Introduction to Biotechnology, 4e, Global Edition (Thieman)

Chapter 3 Recombinant DNA Technology and Genomics

9) Transformation in a cloning experiment is ________.

10) Which of the following is an INCORRECT statement about restriction enzymes?

13) Approximately how large is the human genome?

21) A recombinant DNA molecule produced artificially contains sequences ________.

25) A bacteriophage is ________.

29) Explain the meaning and utility of RNA interference.

30) Explain the dideoxy sequencing method.

31) Which of the following is NOT a feature of a DNA cloning Vector?

39) ________ is a sequencing method that only sequences exons.

You might also like

Pfad - The Proxy pFad of © 2024 Garber Painting. All rights reserved.