8. Figure 20.2
Bacterium
Bacterial
chromosome
Plasmid
2
1
3
4
Gene inserted into
plasmid
Cell containing gene
of interest
Recombinant
DNA (plasmid)
Gene of
interest
Plasmid put into
bacterial cell
DNA of
chromosome
(“foreign” DNA)
Recombinant
bacterium
Host cell grown in culture to
form a clone of cells containing
the “cloned” gene of interest
Gene of
interest
Protein expressed from
gene of interest
Protein harvestedCopies of gene
Basic research
and various
applications
Basic
research
on protein
Basic
research
on gene
Gene for pest
resistance inserted
into plants
Gene used to alter
bacteria for cleaning
up toxic waste
Protein dissolves
blood clots in heart
attack therapy
Human growth
hormone treats
stunted growth
9. Figure 20.2a
Bacterium
Bacterial
chromosome
Plasmid
2
1 Gene inserted into
plasmid
Cell containing
gene of interest
Recombinant
DNA (plasmid)
Gene of
interest
Plasmid put into
bacterial cell
DNA of
chromosome
(“foreign” DNA)
Recombinant
bacterium
10. Figure 20.2b
Host cell grown in
culture to form a clone
of cells containing the
“cloned” gene of interest
Gene of
interest
Protein expressed from
gene of interest
Protein harvestedCopies of gene
Basic research
and various
applications
3
4
Basic
research
on protein
Basic
research
on gene
Gene for pest
resistance inserted
into plants
Gene used to alter
bacteria for cleaning
up toxic waste
Protein dissolves
blood clots in heart
attack therapy
Human growth
hormone treats
stunted growth
13. Figure 20.3-1
Restriction enzyme
cuts sugar-phosphate
backbones.
Restriction site
DNA
5′
5′
5′
5′
5′
5′
3′
3′
3′
3′
3′
3′
1
Sticky
end
GAATTC
CTTAAG
CTTAA
G AATTC
G
14. Figure 20.3-2
One possible combination
DNA fragment added
from another molecule
cut by same enzyme.
Base pairing occurs.
Restriction enzyme
cuts sugar-phosphate
backbones.
Restriction site
DNA
5′
5′
5′
5′
5′
5′
5′
5′
5′5′
5′
5′
5′5′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
2
1
Sticky
end
GAATTC
CTTAAG
CTTAA
G AATTC
G
GG
AATTC
CTTAA
G
G
G
G
AATT CAATT C
C TTAA C TTAA
15. Figure 20.3-3
Recombinant DNA molecule
One possible combination
DNA ligase
seals strands
DNA fragment added
from another molecule
cut by same enzyme.
Base pairing occurs.
Restriction enzyme
cuts sugar-phosphate
backbones.
Restriction site
DNA
5′
5′
5′
5′
5′
5′
5′
5′
5′5′
5′
5′
5′5′
5′
5′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
3′
2
3
1
Sticky
end
GAATTC
CTTAAG
CTTAA
G AATTC
G
GG
AATTC
CTTAA
G
G
G
G
AATT CAATT C
C TTAA C TTAA
25. Figure 20.5
Foreign genome
Cut with restriction enzymes into either
small
fragments
large
fragments
or
Recombinant
plasmids
Plasmid
clone
(a) Plasmid library
(b) BAC clone
Bacterial artificial
chromosome (BAC)
Large
insert
with
many
genes
(c) Storing genome libraries
30. Figure 20.6-2
DNA in
nucleus
mRNAs in
cytoplasm
mRNA
Reverse
transcriptase Poly-A tail
DNA
strand
Primer
5′
5′
3′
3′
A A A A A A
T T T T T
31. Figure 20.6-3
DNA in
nucleus
mRNAs in
cytoplasm
mRNA
Reverse
transcriptase Poly-A tail
DNA
strand
Primer
5′
5′
5′
5′
3′
3′
3′
3′
A A A A A A
A A A A A A
T T T T T
T T T T T
32. Figure 20.6-4
DNA in
nucleus
mRNAs in
cytoplasm
mRNA
Reverse
transcriptase Poly-A tail
DNA
strand
Primer
DNA
polymerase
5′
5′
5′
5′
5′
5′
3′
3′
3′
3′
3′
3′
A A A A A A
A A A A A A
T T T T T
T T T T T
33. Figure 20.6-5
DNA in
nucleus
mRNAs in
cytoplasm
mRNA
Reverse
transcriptase Poly-A tail
DNA
strand
Primer
DNA
polymerase
cDNA
5′
5′
5′
5′
5′
5′
5′
5′
3′
3′
3′
3′
3′
3′
3′
3′
A A A A A A
A A A A A A
T T T T T
T T T T T
37. Figure 20.7
Radioactively
labeled probe
molecules Gene of
interest
Probe
DNA
Single-
stranded
DNA from
cell
Film
Location of
DNA with the
complementary
sequence
Nylon
membrane
Nylon membrane
Multiwell plates
holding library
clones
TECHNIQUE 5′
5′3′
3′
GAGTAGTGGCCG
⋅⋅⋅ CTCATCACCGGC⋅⋅⋅
51. Figure 20.9
Mixture of
DNA mol-
ecules of
different
sizes
Power
source
Power
source
Longer
molecules
Cathode Anode
Wells
Gel
Shorter
molecules
TECHNIQUE
RESULTS
1
2
− +
− +
52. Figure 20.9a
Mixture of
DNA mol-
ecules of
different
sizes
Power
source
Power
source
Longer
molecules
Cathode Anode
Wells
Gel
Shorter
molecules
TECHNIQUE
2
− +
− +
1
56. Figure 20.10
Normal β-globin allele
Sickle-cell mutant β-globin allele
Large
fragment
Normal
allele
Sickle-cell
allele
201 bp
175 bp
376 bp
(a) DdeI restriction sites in normal and
sickle-cell alleles of the β-globin gene
(b)Electrophoresis of restriction
fragments from normal and
sickle-cell alleles
201 bp175 bp
376 bp
Large fragment
Large fragment
DdeI DdeI DdeI DdeI
DdeI DdeI DdeI
57. Figure 20.10a
Normal β-globin allele
Sickle-cell mutant β-globin allele
(a) DdeI restriction sites in normal and
sickle-cell alleles of the β-globin gene
201 bp175 bp
376 bp
Large fragment
Large fragment
DdeI DdeI DdeI DdeI
DdeI DdeI DdeI
60. Figure 20.11
DNA + restriction enzyme
321
4
TECHNIQUE
I Normal
β-globin
allele
II Sickle-cell
allele
III Heterozygote
Restriction
fragments
Nitrocellulose
membrane (blot)
Heavy
weight
Gel
Sponge
Alkaline
solution Paper
towels
III III
III III III III
Preparation of
restriction fragments
Gel electrophoresis DNA transfer (blotting)
Radioactively labeled
probe for β-globin
gene
Nitrocellulose blot
Probe base-pairs
with fragments
Fragment from
sickle-cell
β-globin allele
Fragment from
normal β- globin
allele
Film
over
blot
Hybridization with labeled probe Probe detection5
62. Figure 20.12
DNA
(template strand)
TECHNIQUE
5′
3′
C
C
C
C
T
T
T
G
G
A
A
A
A
G
T
T
T
DNA
polymerase
Primer
5′
3′
P P P
OH
G
dATP
dCTP
dTTP
dGTP
Deoxyribonucleotides Dideoxyribonucleotides
(fluorescently tagged)
P P P
H
G
ddATP
ddCTP
ddTTP
ddGTP
5′
3′
C
C
C
C
T
T
T
G
G
A
A
A
A
DNA (template
strand)
Labeled strands
Shortest Longest
5′
3′
ddC
ddG
ddA
ddA
ddA
ddG
ddG
ddT
ddC
G
T
T
T
G
T
T
T
C
G
T
T
T
C
T T
G
G
T
T
T
C
T
G
A
G
T
T
T
C
T
G
A
A
G
T
T
T
C
T
G
A
A
G
G
T
T
T
C
T
G
A
A
G
T
G
T
T
T
C
T
G
A
A
G
T
C
G
T
T
T
C
T
G
A
A
G
T
C
A
Direction
of movement
of strands
Longest labeled strand
Detector
Laser
Shortest labeled strand
RESULTS
Last nucleotide
of longest
labeled strand
Last nucleotide
of shortest
labeled strand
G
G
G
A
A
A
C
C
T
63. Figure 20.12a
DNA
(template strand)
TECHNIQUE
Primer Deoxyribonucleotides Dideoxyribonucleotides
(fluorescently tagged)
DNA
polymerase
5′
5′
3′
3′
OH H
GG
dATP
dCTP
dTTP
dGTP
P P P P P P
ddATP
ddCTP
ddTTP
ddGTP
T
T
T
G
G
G
C
C
C
C
T
T
T
A
A
A
A
64. Figure 20.12b
DNA (template
strand)
Labeled strands
Shortest Longest
Direction
of movement
of strands
Longest labeled strand
Detector
Laser
Shortest labeled strand
TECHNIQUE (continued)
5′
3′
G
G
C
C
C
C
T
T
T
A
A
A
A
T
T
T
G
ddC
ddC
ddG
ddG
ddG
ddA
ddA
ddA
ddT
3′
5′
T
T
T
G
C
T
T
T
G
C
G
T
T
T
G
C
G
A
T
T
T
G
C
G
A
A
T
T
T
G
C
G
A
A
G
T
T
T
C
G
A
A
G
T
T
T
T
C
G
A
A
G
T
C
A
T
T
T
C
G
A
A
G
T
C
G G G
65. Figure 20.12c
RESULTS
Last nucleotide
of longest
labeled strand
Last nucleotide
of shortest
labeled strand
G
G
G
A
A
A
C
C
T
Direction
of movement
of strands
Longest labeled strand
Detector
Laser
Shortest labeled strand
74. Isolate mRNA.
2
1
3
4
TECHNIQUE
Make cDNA by reverse
transcription, using
fluorescently labeled
nucleotides.
Apply the cDNA mixture to a
microarray, a different gene
in each spot. The cDNA hybridizes
with any complementary DNA on
the microarray.
Rinse off excess cDNA; scan microarray
for fluorescence. Each fluorescent spot
(yellow) represents a gene expressed
in the tissue sample.
Tissue sample
mRNA molecules
Labeled cDNA molecules
(single strands)
DNA fragments
representing a
specific gene
DNA microarray
DNA microarray
with 2,400
human genes
Figure 20.15
82. Figure 20.17
Cross
section of
carrot root
2-mg
fragments
Fragments were
cultured in nu-
trient medium;
stirring caused
single cells to
shear off into
the liquid.
Single cells
free in
suspension
began to
divide.
Embryonic
plant developed
from a cultured
single cell.
Plantlet was
cultured on
agar medium.
Later it was
planted in soil.
Adult
plant
88. 4
5
6
RESULTS
Grown in culture
Implanted in uterus
of a third sheep
Embryonic
development
Nucleus from
mammary cell
Early embryo
Surrogate
mother
Lamb (“Dolly”) genetically
identical to mammary cell donor
Figure 20.19b
95. Figure 20.22
Remove skin cells
from patient. 2
1
3
4
Reprogram skin cells
so the cells become
induced pluripotent
stem (iPS) cells.
Patient with
damaged heart
tissue or other
disease
Return cells to
patient, where
they can repair
damaged tissue.
Treat iPS cells so
that they differentiate
into a specific
cell type.
100. Figure 20.23
Cloned gene
2
1
3
4
Retrovirus
capsid
Bone
marrow
cell from
patient
Viral RNA
Bone
marrow
Insert RNA version of normal allele
into retrovirus.
Let retrovirus infect bone marrow cells
that have been removed from the
patient and cultured.
Viral DNA carrying the normal
allele inserts into chromosome.
Inject engineered
cells into patient.
110. Figure 20.25
This photo shows
Washington just before
his release in 2001,
after 17 years in prison.
(a)
(b)These and other STR data exonerated Washington
and led Tinsley to plead guilty to the murder.
Semen on victim
Earl Washington
Kenneth Tinsley
17,19
16,18
17,19
13,16
14,15
13,16
12,12
11,12
12,12
Source of
sample
STR
marker 1
STR
marker 2
STR
marker 3
111. Figure 20.25a
This photo shows
Washington just before
his release in 2001,
after 17 years in prison.
(a)
115. Figure 20.26
Plant with new trait
RESULTS
TECHNIQUE
Ti
plasmid
Site where
restriction
enzyme cuts
DNA with
the gene
of interest
Recombinant
Ti plasmid
T DNA
Agrobacterium tumefaciens