Chapter 12
DNA Technology and
Genomics
PowerPoint TextEdit Art Slides for
Biology: Concepts and Connections, Fifth Edition
– Campbell, Reece, Taylor, and Simon
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.1 An overview of gene cloning (layer 1)
Bacterium
1
Plasmid
isolated
Cell containing gene
of interest
2
3
Bacterial
chromosome
Plasmid
DNA
isolated
Gene inserted
into plasmid
Gene of
interest
Recombinant DNA
(plasmid)
4
DNA
Plasmid put into
bacterial cell
Recombinant
bacterium
5
Cell multiplies with
gene of interest
Copies of protein
Copies of gene
Clone of cells
Gene for pest
resistance
inserted into
plants
Gene used to alter bacteria
for cleaning up toxic waste
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Protein used to
make snow form
at higher
temperature
Protein used to dissolve blood
clots in heart attack therapy
Figure 12.1 An overview of gene cloning (layer 2)
Bacterium
1
Plasmid
isolated
Cell containing gene
of interest
2
3
Bacterial
chromosome
Plasmid
DNA
isolated
Gene inserted
into plasmid
Gene of
interest
Recombinant DNA
(plasmid)
4
DNA
Plasmid put into
bacterial cell
Recombinant
bacterium
5
Cell multiplies with
gene of interest
Copies of protein
Copies of gene
Clone of cells
Gene for pest
resistance
inserted into
plants
Gene used to alter bacteria
for cleaning up toxic waste
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Protein used to
make snow form
at higher
temperature
Protein used to dissolve blood
clots in heart attack therapy
Figure 12.1 An overview of gene cloning (layer 3)
Bacterium
1
Plasmid
isolated
Cell containing gene
of interest
2
3
Bacterial
chromosome
Plasmid
DNA
isolated
Gene inserted
into plasmid
Gene of
interest
Recombinant DNA
(plasmid)
4
DNA
Plasmid put into
bacterial cell
Recombinant
bacterium
5
Cell multiplies with
gene of interest
Copies of protein
Copies of gene
Clone of cells
Gene for pest
resistance
inserted into
plants
Gene used to alter bacteria
for cleaning up toxic waste
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Protein used to
make snow form
at higher
temperature
Protein used to dissolve blood
clots in heart attack therapy
Figure 12.2 Creating recombinant DNA using a
restriction enzyme and DNA ligase
Restriction enzyme
recognition sequence
1
GAATTC
CTTAAG
DNA
Restriction enzyme
cuts the DNA into
fragments
2
Sticky end
Addition of a DNA
fragment from
another source
A AT TC
G
3
Two (or more)
fragments stick
together by
base-pairing
4
G A AT T C
C T TA A G
G A AT T C
C T TA A G
DNA ligase
Pastes the strand
5
Recombinant DNA molecule
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.3 Cloning a gene in a bacterial plasmid (layer 1)
Human cell
E.coli
1 Isolate DNA
from two sources
2 Cut both DNAs
with the same
restriction enzyme
DNA
Plasmid
Gene V
Sticky ends
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.3 Cloning a gene in a bacterial plasmid (layer 2)
Human cell
E.coli
1 Isolate DNA
from two sources
2 Cut both DNAs
with the same
restriction enzyme
DNA
Plasmid
Gene V
Sticky ends
3 Mix the DNAs;
they join by
base-pairing
4 Add DNA ligase
to bond the DNA covalently
Recombinant DNA
plasmid
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Gene V
Figure 12.3 Cloning a gene in a bacterial plasmid (layer 3)
Human cell
E.coli
1 Isolate DNA
from two sources
2 Cut both DNAs
with the same
restriction enzyme
DNA
Plasmid
Gene V
Sticky ends
3 Mix the DNAs;
they join by
base-pairing
4 Add DNA ligase
to bond the DNA covalently
Recombinant DNA
plasmid
Gene V
5 Put plasmid into bacterium
by transformation
Recombinant
bacterium
6 Clone the bacterium
Bacterial clone carrying many
copies of the human gene
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
12.3 Cloning a gene
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.7A Human insulin produced by bacteria
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.18B Transgenic pigs
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.6 “Pharm” animals that produce a
human protein
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.18A Using the Ti plasmid as a vector for
genetically engineering plants
Agrobacterium tumefaciens
DNA containing
gene for desired trait
Ti
plasmid
1
Insertion of gene
into plasmid using
restriction enzyme
and DNA ligase
T DNA
Recombinant
Ti plasmid
Restriction
site
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Plant cell
2
3
Introduction
Regeneration
into plant
of plant
cells in
culture
Plant with new trait
T DNA carrying new
gene within plant chromosome
Figure 12.19B Pollen might transfer genes from genetically
engineered crop plants to wild relatives nearby
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Table 12.6 Some protein products of recombinant
DNA technology
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.14 DNA amplification by PCR
Initial
DNA
segment
1
2
4
Number of DNA molecules
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
8
Figure 12.11A Restriction site differences between
two homologous samples of DNA
Crime scene
Suspect
w
Cut
C
C
G
G
G
G
C
C
A T
C G
G C
G C
z
x
Cut
y
C
C
G
G
G
G
C
C
Cut
y
C
C
G
G
DNA from chromosomes
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
G
G
C
C
Figure 12.10 Gel electrophoresis of DNA
Mixture of DNA
molecules of
different sizes
–
–
Longer
molecules
Power
source
Gel
+
Shorter
molecules
+
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Completed gel
Figure 12.12A DNA fingerprints from a murder case
Defendant’s
blood
Blood from
defendant’s clothes
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Victim’s
blood
Figure 12.12B DNA data for forensic use
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Unnumbered Figure p. 231
Investigator at one
of the crime scenes
(above), Narborough,
England (left)
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Unnumbered Figure p. 230
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.8 How a DNA probe tags a gene by base pairing
Radioactive
probe (DNA)
Mix with singlestranded DNA from
various bacterial
(or phage) clones
Single-stranded
DNA
Base pairing
indicates the
gene of interest
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.11B Gel electrophoresis of restriction fragments
1
–
2
Longer
fragments
z
x
w
shorter
fragments
y
+
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
y
Figure 12.11C The use of restriction fragment
analysis to detect a harmful allele (layer 1)
1 Restriction fragment
preparation
I
II
III
Restriction
fragments
2 Gel electrophoresis
I II III
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.11C The use of restriction fragment
analysis to detect a harmful allele (layer 2)
1 Restriction fragment
preparation
I
II
III
Restriction
fragments
2 Gel electrophoresis
I II III
3 Blotting
Filter paper
4 Radioactive probe
Radioactive, singlestranded DNA (probe)
Probe
I
II
III
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.11C The use of restriction fragment
analysis to detect a harmful allele (layer 3)
1 Restriction fragment
preparation
I
II
III
Restriction
fragments
2 Gel electrophoresis
I II III
3 Blotting
Filter paper
4 Radioactive probe
Radioactive, singlestranded DNA (probe)
Probe
5 Detection of radioactivity
(autoadiography)
I
II
III
Film
I
II
III
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.7B Equipment used in the production of
a vaccine against hepatitis B
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.13 One type of gene therapy procedure
Cloned gene
(normal allele)
1 Insert normal gene
into virus
Viral nucleic
acid
Retrovirus
2 Infect bone marrow
cell with virus
3 Viral DNA inserts
into chromosome
Bone marrow
cell from patient
Bone
marrow
4 Inject cells
into patient
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.9 DNA microarray
DNA microarray
Each well contains DNA
from a particular gene
1 mRNA
isolated
Reverse transcriptase
and fluorescent DNA
nucleotides
2 cDNA made
from mRNA
Actual size
(6,400 genes)
4 Unbound
cDNA rinsed
away
Fluorescent
spot
3 cDNA applied
to wells
cDNA
DNA of an
expressed gene
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Nonfluorescent
spot
DNA of an
unexpressed gene
Figure 12.15 DNA sequencing
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Table 12.17 Some important completed genomes
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.19A A maximum-security laboratory at
the Pasteur Institute in Paris
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Figure 12.20 Eric Lander
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Unnumbered Figure p. 250
Bacterial
clone
Cut
Bacterium
DNA
fragments
Cut
Recombinant
DNA
plasmids
Recombinant
bacteria
Plasmids
Genomic library
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Unnumbered Figure p. 250
Mixture
of DNA
fragments
–
Longer
fragments
move slower
A “band” is
a collection
of DNA
fragments of
one particular
length
Power
source
Shorter
fragments
move faster
+
DNA attracted to +
pole due to PO4– groups
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Unnumbered Figure p. 250
Unknown
DNA
amplified
via
(a)
Bacterial
plasmids
Unknown
sample
Suspect
sample for
comparison
treated with treated with
treated with
(b)
DNA
fragments
sorted by size via
Visualizing and
comparing
fragments is
a method called
(c)
Recombinant plasmids
are inserted
into bacteria
(d)
Add
(e)
Particular
DNA
sequence
highlighted
(g)
Collection
is called a
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
are copied via
(f)
Figure 12.4 Genomic libraries
Genome cut up with
restriction enzyme
Recombinant
plasmid
Recombinant
phage DNA
or
Bacterial
clone
Plasmid library
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings
Phage
clone
Phage library
Figure 12.5 Making an intron-lacking gene from
eukaryotic mRNA
Cell nucleus
Exon Intron
Exon
Intron Exon
DNA of
eukaryotic
gene
1 Transcription
RNA
transcript
2 RNA splicing
(removes introns)
mRNA
Test tube
Reverse transcriptase
3 Isolation of mRNA
from cell and addition
of reverse transcriptase;
synthesis of DNA strand
cDNA strand
4 Breakdown of RNA
5 Synthesis of second
DNA strand
cDNA of gene
(no introns)
Copyright © 2005 Pearson Education, Inc. Publishing as Benjamin Cummings