Chapter 20

DNA Technology
and Genomics
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Biology, Seventh Edition
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Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings
 Overview: Understanding and Manipulating
Genomes
One of the greatest achievements of modern
science
Has been the sequencing of the human
genome, which was largely complete by 2003

DNA sequencing accomplishments

Have all depended on advances in DNA
technology, starting with the invention of
methods for making recombinant DNA
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 Concept 20.1: DNA cloning permits production
of multiple copies of a specific gene or other
DNA segment
To work directly with specific genes
Scientists have developed methods for
preparing well-defined, gene-sized pieces of
DNA in multiple identical copies, a process
called gene cloning

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 DNA Cloning and Its Applications: A Preview
Most methods for cloning pieces of DNA in the
laboratory
Share certain general features, such as the
use of bacteria and their plasmids

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 Overview of gene cloning with a bacterial
plasmid, showing various uses of cloned genes
Bacterium Cell containing gene
1 Gene inserted of interest
into plasmid

Bacterial Plasmid Gene of

chromosome interest
Recombinant DNA of
DNA (plasmid) 2 Plasmid put into chromosome
bacterial cell

Recombinate
bacterium
3 Host cell grown in culture,
to form a clone of cells
containing the cloned
gene of interest
Gene of
Protein expressed
interest
by gene of interest
Copies of gene Protein harvested

Basic 4 Basic research and Basic

research various applications research
on protein
Universality of the on gene

genetic code
enables this progress
Gene for pest Gene used to alter Protein dissolves Human growth
resistance inserted bacteria for cleaning blood clots in heart hormone treats
Figure 20.2 into plants up toxic waste attack therapy stunted growth

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 Using Restriction Enzymes to Make Recombinant DNA
Bacterial restriction enzymes
Cut DNA molecules at a limited number of
specific DNA sequences, called restriction
sites

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 A restriction enzyme will usually make many
cuts in a DNA molecule
Yielding a set of restriction fragments
The most useful restriction enzymes cut DNA
in a staggered way
Producing fragments with sticky ends that
can bond with complementary sticky ends of
other fragments
DNA ligase is an enzyme
That seals the bonds between restriction
fragments
Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings
 Using a restriction enzyme and DNA ligase to
make recombinant DNA
Restriction site

DNA 5 GAATTC 3
3 CTTAAG 5

1 Restriction enzyme cuts

the sugar-phosphate
backbones at each arrow

G AATTC
CTTAA G

Sticky end
AATTC
G
G
2 DNA fragment from CTTAA

another source is added. Fragment from different

Base pairing of sticky DNA molecule cut by the
ends produces various same restriction enzyme
combinations.
G AATT C G AATTC
C TTAA G CTTAA G

One possible combination

3 DNA ligase
seals the strands.

Figure 20.3 Recombinant DNA molecule

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 Cloning a Eukraryotic Gene in a Bacterial Plasmid
In gene cloning, the original plasmid is called a
cloning vector
Defined as a DNA molecule that can carry
foreign DNA into a cell and replicate there

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 Producing Clones of Cells
APPLICATION Cloning is used to prepare many copies of a gene of interest for use in sequencing the gene,
in producing its encoded protein, in gene therapy, or in basic research.

TECHNIQUE In this example, a human gene is inserted into a plasmid from E. coli. The plasmid contains
the ampR gene, which makes E. coli cells resistant to the antibiotic ampicillin. It also contains
the lacZ gene, which encodes -galactosidase. This enzyme hydrolyzes a molecular mimic of
lactose (X-gal) to form a blue product. Only three plasmids and three human DNA fragments
are shown, but millions of copies of the plasmid and a mixture of millions of different human
DNA fragments would be present in the samples.

Bacterial cell lacZ gene

1 Isolate plasmid DNA and human DNA. (lactose Human
breakdown) cell

Restriction
site
2 Cut both DNA samples with the same restriction
ampR gene
enzyme (ampicillin Bacterial Gene of
plasmid interest
resistance)

Sticky
ends Human DNA
3 Mix the DNAs; they join by base pairing. fragments
The products are recombinant plasmids and
many nonrecombinant plasmids.

Figure 20.4 Recombinant DNA plasmids

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 Concept 20.5: The practical applications of
DNA technology affect our lives in many ways
Numerous fields are benefiting from DNA
technology and genetic engineering

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 Medical Applications
One obvious benefit of DNA technology
Is the identification of human genes whose
mutation plays a role in genetic diseases

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 Diagnosis of Diseases
Medical scientists can now diagnose hundreds
of human genetic disorders
By using PCR and primers corresponding to
cloned disease genes, then sequencing the
amplified product to look for the disease-
causing mutation

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

 Human Gene Therapy
Gene therapy
Is the alteration of an afflicted individuals
genes
Holds great potential for treating disorders
traceable to a single defective gene
Uses various vectors for delivery of genes into
cells

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings

 Gene therapy using a retroviral vector
Cloned gene
(normal
allele, 1 Insert RNA version of normal allele
absent into retrovirus.
from
patients
cells) Viral RNA

2 Let retrovirus infect bone marrow cells

that have been removed from the
Retrovirus patient and cultured.
capsid

3 Viral DNA carrying the normal

allele inserts into chromosome.

Bone
marrow
cell from
patient

4 Inject engineered
Figure 20.16 cells into patient.

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 Pharmaceutical Products
Applications of DNA technology include
Large-scale production of human hormones
and other proteins with therapeutic uses
Production of safer vaccines

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 Forensic Evidence
DNA fingerprints obtained by analysis of
tissue or body fluids found at crime scenes
Can provide definitive evidence that a suspect
is guilty or not
Can a woman leave DNA evidence at a crime
scene?

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 Environmental Cleanup
Genetic engineering can be used to modify the
metabolism of microorganisms
So that they can be used to extract minerals
from the environment or degrade various types
of potentially toxic waste materials

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 Agricultural Applications
DNA technology
Is being used to improve agricultural
productivity and food quality

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 Animal Husbandry and Pharm Animals
Transgenic animals
Contain genes from other organisms

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 Have been engineered to be pharmaceutical
factories

Figure 20.18
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 Genetic Engineering in Plants
Agricultural scientists
Have already endowed a number of crop
plants with genes for desirable traits

Copyright 2005 Pearson Education, Inc. publishing as Benjamin Cummings