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Genes, Genomes

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Title: Genes, Genomes


1
Genes, Genomes Genetic Engineering
Part 2
  • Dan Graur
  • University of Houston

http//nsm.uh.edu/dgraur/
2
Genetic Engineering A collection of techniques
used to manipulate genes, genomes, and other
pieces of DNA, as well as their products and
expression.
3
1. Isolating DNA
4
2. Reading DNA
sequencing
5
3. Cutting DNA
6
Restriction Endonucleases
Protect bacteria from bacteriophages.
7
Restriction Endonucleases
Palindromes Madam, Im Adam. RADAR A dog. A
panic in a pagoda. Campus motto Bottoms up, Mac.
8
Molecular Palindrome
9
4. Ligating DNA
10
ligase
Fragments that have been produced by a
restriction enzyme can be ligated to other
fragments produced by the same enzyme.
11
5. Separation by size
12
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13
6. Identification
14
7. Transportation
15
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16
Texas-style molecular biology
17
8. Selection
18
Plasmid pBR322
19
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20
1
2
6
7
3
4
8
5
Ampicillin
Tetracycline
1 2 3 4
1 2 3 4
Streak individual colonies with sterile toothpick.
5 6 7 8
5 6 7 8
1 2 3 4
1 2 3 4
Incubate overnight.
5 6 7 8
5 6 7 8
21
1
2
6
7
3
4
8
5
Q Which colony is resistant to ampicillin and
sensitive to tetracycline. A 6
Ampicillin
Tetracycline
1 2 3 4
1 2 3 4
5 6 7 8
5 6 7 8
1 2 3 4
1 2 3 4
5 6 7 8
5 6 7 8
22
9. Reverse transcription
23
10. Copy
24
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25
Cycle 0 1 2 3 4 5 n Number 1 2 4 8 16 64 2n of
copies
26
11. Genomic library
Library
27
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28
27 February 1997
12. Cloning
29
Dolly
Incubator
30
Another Dolly
31
13 Many Present Techniques I Know Nothing About
Many Future Techniques
32
Engineering is artificial. There is nothing
artificial about genetic engineering.
33
Oocyte from Drosophila infected with Wolbachia.
Note concentration of bacteria in the anterior
region of the cell.
Drosophila ananassae
D. ananassae is a natural transgenic
34
What is genetic engineering good for?
35
1. Diagnostics of genetic traits (e.g.,
disorders)
36
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37
2. Protein Synthesis
38
Coagulation Factors
39
(1) Vessel damage (2) Vascular spasm (3)
Platelet plug forms. (4) Coagulation.
40
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41
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42
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43
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44
Hemophilias
45
Treating hemophilia A and hemophilia B Factors
VIII and IX are purified from the pooled blood of
several thousands donors. Injections of factors
VIII and IX have allowed hemophiliacs to live
relatively normal lives.
1/2
46
Treating hemophilia A and hemophilia B In the
1980s, HIV-contaminated blood was unknowingly
used to manufacture factors VIII and IX. In
some countries (e.g., France), gt 90 of the
hemophiliacs became infected by HIV.
2/2
47
Possibility of HIV contamination
48
Human Growth Hormone The first GE product.
49
Is Jintropin (recombinant human growth hormone)
the new national past time?
50
Insulin The best selling GE product.
51
3. Transgenics
52
Photinus pyralis A source of luciferase
53
Insert luciferase gene into tobacco cell culture,
cause differentiation to occur, water plants with
ATP solution, and get a glowing-in-the-dark
transgenic tobacco plant.
54
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55
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56
Green fluorescent protein (GFP)
57
Attach to Opsin promoter Expression in Eyes
Attach to Lysin promoter Expression in Sperm
58
4. Personal identification (Fingerprinting)
59
People are different from one another
60
Paternity determination
61
Paternity determination
62
How to distinguish between a relatively cheap
Trebbiano and a much more expensive Verdelho?
63
expensive
cheap
64
5. Somatic gene therapy
65
Injection of Transformed Cells
66
Severe combined immunodeficiency (SCID) due to
adenosine deaminase (ADA) deficiency
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