Professional Documents
Culture Documents
p.174
Biotechnologyuseoflivingorganismstocreateproducts
orhelpprocesses
Ex.HGH,insulin
RecombinantDNAsegmentofDNAcontaining
sequencesfromdifferentorganisms
HowisDNAmanipulated?
RestrictionenzymescutDNAatspecificsites
andcreatestickyends
G T G G
T
T
A A
A C C
C
A
A
T T
T A
C C T
C C A
G A
G G A
G G T C
T
G A
T C
G
T
G
C T C
T
T
C A G
A
A
A
C
A
C
T
G
G A
C A
A
G G
T C T
T T A
GA
GA
T
TC C
C
G
T
T G
C T
T
C G A
T T G
A
A
C C A G AA C G G
A
T TC
T
C
C
T
A
A
G
C
C
A
G
G
G
A
C
A
A
G
A
T
A
C
G G T C T T G G C
TA A G
T T
Complementaryendswillfusetoproduce
alongstrandofDNA
Degraded DNA
Crossovers
Recipient cells
chromosome
Figure 12.1D
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings
Recombinant
chromosome
PlasmidsareextraringsofDNAthatreplicateinbacteria.
DNAcanbeinsertedintoplasmids.
bacterium
bacterial
chromosome
plasmid
CloningVectors
Bacterium
Human cell
Plasmid
DNA
Human protein
Bacterial
chromosome
recombinant DNA
transformation
The polymerase
chain reaction
(PCR) can
quickly clone a
small sample of
DNA in a test
tube
Initial
DNA
segment
Selection of
specific sequence
1
8
Figure 12.12
Longer
molecules
Power
source
Gel
Shorter
molecules
Glass
plates
Completed gel
Figure 12.10
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings
DNA
forensics
Eggmicroinjectiontoproduce
transgenicanimal
Growbiggerfishfaster.
Salmonwithgenefromanotherfishspecies
Uses of
transformed
animals:
Produce
medicines more
easily
Ex. sheep and gene to
treat cystic fibrosis
Goats and AT3 gene to
prevent blood clots
Figure 12.16
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings
Geneticengineeringofplants
MethodstoinsertDNA:
1. Ballistics
2. Protoplasts
3. Agrobacteriumasvector
Tiplasmid
a A bacterial
cell contains
a Ti plasmid
(purple) that
has a foreign
gene (blue).
b The bacterium
infects a plant
and transfers the
Ti plasmid into it.
The plasmid DNA
becomes
integrated into
one of the plants
chromosomes.
A young
plant
expressing
a fluorescent
gene product
Benefitsandrisks
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings
HowDollywascloned
DNA
udder cells
white sheep
embryo
surrogate
mother
egg cell
black sheep
Dolly
Cloningofhumancells
Regenerativemedicine
Bone,pancreascells,skin
Stemcellsthe$6billionpromise?
normal gene
into virus
Viral nucleic
acid
Ex vivo
Retrovirus
In vivo
2 Infect bone
marrow cell
with virus
Stem cells
3 Viral DNA
inserts into
chromosome
Bone marrow
cell from patient
Bone
marrow
Figure 12.19
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings
4 Inject cells
into patient
HumanGenomeProject
3.2billionbasesin22autosomes+X,Y
Draftsequencecompletedin2003
Availableat
www.ornl.gov/sci/techresources/Human_Genome/
home.shtml
www.ucsc.genome.edu
Whatdoesthehumangenome
sequencetellus?
20Kto25Kgenes
99.9%alike,acrossallraces
97%ofDNAisnottranscribed
Spacersbetweengenes
Structural(centromeres,telomeres)
Regulatory(enhancers,promoters)
Leftoversofevolution?
Howarespecificgenesidentified?
1.Isolateitfromagenomiclibraryby
homologywithagenefromanother
organism.
2. FindmRNAforthegene,makecDNA
fromit.
3. MakeDNAsequencebasedonprotein
sequence.
Base pairing
indicates the
gene of interest
Figure 12.8A
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings
cDNA
mRNA
Complementary
DNA
mRNA
Using reverse
cDNA
transcriptase
Assembles DNA
on mRNA
template
reverse
transcriptase
DNA
polymerase
DNA
DNA
Fig. 11-4, p.164
cDNA
DNA of gene
DNA
microarray,
actual size
(6,400 genes)
Figure 12.9
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings
Gene Therapy
What is it?
How is it done?
Does it work?
Gene therapy
Goal - Treat diseases caused by mutated genes
Method - Add a normal gene or block an
abnormal gene in enough cells to restore
normal function
Target - somatic cells
1.
2.
3.
4.
5.
Gene size
Target tissue