Chapter 14: Genetic Engineering -Modification of the DNA of an organism to produce new genes with new characteristics - PowerPoint PPT Presentation

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Chapter 14: Genetic Engineering -Modification of the DNA of an organism to produce new genes with new characteristics

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Title: Chapter 14: Genetic Engineering -Modification of the DNA of an organism to produce new genes with new characteristics


1
Chapter 14 Genetic Engineering-Modification of
the DNA of an organism to produce new genes with
new characteristics
2
Biotechnology
  • Use of organisms to benefit humanity

3
Recombinant DNA technology
  • DNA from different organisms is spliced together
  • Allows scientists to make many copies of any DNA
    segment (clone)
  • Can introduce foreign DNA into cells of
    microorganisms

4
Recombinant DNA technology
  • Restriction enzymes cut DNA
  • Bacteria produce for defense against viruses
  • Vector transports DNA into a cell
  • Ex bacteriophage
  • Plasmid separate, smaller circular DNA that
    maybe be present and able to replicate inside
    bacteria
  • Transformation uptake of foreign DNA by cells
  • How plasmids can get into bacteria

5
Bacterial Conjugation and Recombination
6
Recombinant DNA technology
  • Palindromic sequences reads the same as
    complement, in opposite direction
  • AAGCTT
  • TTCGAA
  • Many restriction enzymes cut these sequences
  • Restriction enzymes cut on a stagger ? sticky
    ends (can pair with complementary single-stranded
    end of other DNA cut with same enzyme)
  • DNA Ligase links 2 fragments ? recombinant DNA

7
Fig. 20-3-3
Restriction site
5? 3?
3? 5?
DNA
Restriction enzymecuts sugar-phosphatebackbones.
1
Sticky end
DNA fragment addedfrom another moleculecut by
same enzyme.Base pairing occurs.
2
One possible combination
DNA ligaseseals strands.
3
Recombinant DNA molecule
8
Restriction Enzymes
9
Steps of Creating a Recombinant DNA Plasmid
(Basic)
  • 1. Plasmids and desired DNA cut by same
    restriction enzyme
  • 2. Mix 2 types of DNA so sticky ends pair
  • 3. DNA ligase forms bonds between fragments

10
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11
Fig. 20-2
Cell containing geneof interest
Bacterium
1
Gene inserted intoplasmid
Bacterialchromosome
Plasmid
Gene ofinterest
RecombinantDNA (plasmid)
DNA of chromosome
2
Plasmid put intobacterial cell
Recombinantbacterium
Host cell grown in cultureto form a clone of
cellscontaining the clonedgene of interest
3
Gene ofInterest
Protein expressedby gene of interest
Copies of gene
Protein harvested
Basic research andvarious applications
4
Basicresearchon protein
Basicresearchon gene
Gene for pest resistance inserted into plants
Gene used to alter bacteria for cleaning up
toxic waste
Protein dissolvesblood clots in heartattack
therapy
Human growth hor-mone treats stuntedgrowth
12
Cloning a Gene
13
Fig. 20-4-4
Hummingbird cell
TECHNIQUE
Bacterial cell
lacZ gene
Restrictionsite
Gene of interest
Stickyends
Bacterial plasmid
ampR gene
Hummingbird DNA fragments
Nonrecombinant plasmid
Recombinant plasmids
Bacteria carryingplasmids
RESULTS
Colony carrying recombinant plasmid with
disrupted lacZ gene
Colony carrying non-recombinant plasmidwith
intact lacZ gene
One of manybacterial clones
14
Cloning DNA
  • Genome total DNA per cell
  • Genomic library collection of DNA fragments
    more or less representative of all DNA in genome
  • Genetic Probe single stranded DNA or RNA that
    is radioactively labeled and can attach to target
    sequence by base pairing rules

15
  • A probe can be synthesized that is complementary
    to the gene of interest
  • For example, if the desired gene is
  • Then we would synthesize this probe



5?
G
3?
G
G
C
C
C
T
T
T
A
A
A
C
3?
5?
C
C
G
G
G
A
A
A
T
T
T
16
DNA Probe
17
Using a DNA probe
18
Polymerase Chain Reaction (PCR)
  • Can amplify a small sample of DNA quickly
  • DNA replication in vitro
  • 2 strands separated by heating so special
    heat-resistant DNA polymerase called Taq
    polymerase used (thermophile)
  • MAJOR BONUS Only specific sequences can be
    replicated
  • Study crime scenes, archaeological remains

19
PCR
20
Gel Electrophoresis
  • Separates fragments like DNA, RNA or polypeptides
    (they carry charge and can migrate in an
    electrical field
  • RNA and DNA (-) --- so they move to () pole
  • Smaller fragments go further
  • Compare sample to standard
  • Usually blot - transfer DNA from gel to
    nitrocellulose filter for further analysis
  • DNA Fingerprinting

21
Gel Electrophoresis
22
Fig. 20-9a
TECHNIQUE
Powersource
Mixture ofDNA mol-ecules ofdifferentsizes
Anode
Cathode


Gel
1
Powersource


Longermolecules
2
Shortermolecules
23
Fig. 20-9b
RESULTS
24
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25
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26
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27
DNA Fingerprint
28
Transgenic Organisms
  • Plants and animals in which foreign genes have
    been incorporated
  • Animals
  • Inject DNA into nucleus of egg or stem cell
  • Eggs implanted in uterus stem cells injected
    into blastocysts then implanted into foster
    mother
  • Plants
  • Disease resistance
  • Pesticide resistance

29
Transgenics
30
Fig. 20-20
Adult stem cells
Embryonic stem cells
Early human embryoat blastocyst stage(mammalian
equiva-lent of blastula)
From bone marrowin this example
Cells generatingall embryoniccell types
Cells generatingsome cell types
Culturedstem cells
Differentcultureconditions
Differenttypes ofdifferentiatedcells
Blood cells
Nerve cells
Liver cells
31
Fig. 20-18
TECHNIQUE
Mammarycell donor
Egg celldonor
1
2
Egg cellfrom ovary
Nucleusremoved
Cells fused
Culturedmammary cells
3
3
Nucleus frommammary cell
Grown inculture
4
Early embryo
Implantedin uterusof a thirdsheep
5
Surrogatemother
Embryonicdevelopment
6
Lamb (Dolly)genetically identical tomammary
cell donor
RESULTS
32
Fig. 20-19
33
Cloning Video
34
GE Plants
35
Application of GE
  • Human proteins
  • Insulin
  • Hormones - HGH
  • Human treatments for disease
  • Multiple sclerosis, certain cancers, heart
    attacks, forms of anemia
  • Vaccines

36
Fig. 20-23
37
Fig. 20-24
(a)
This photo shows EarlWashington just before his
release in 2001,after 17 years in prison.
Source of sample
STRmarker 1
STRmarker 2
STRmarker 3
Semen on victim
17, 19
13, 16
12, 12
Earl Washington
16, 18
14, 15
11, 12
17, 19
13, 16
12, 12
Kenneth Tinsley
(b)
These and other STR data exonerated Washington
andled Tinsley to plead guilty to the murder.
38
Forensics
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