Chapter 13 Biotechnology Techniques

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Chapter 13 Biotechnology Techniques
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• Chapter 13.
• 1. Purifying and detecting nucleic acids
• Cloning and recombinant DNA
• Genetic engineering.
• DNA libraries.

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• Purifying and detecting nucleic acids

Separaation of nucleic acids Electrophoresis
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Detection of Nucleic Acids Radioactivity 32P
or 35S Fluoresce light emission
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2. Cloning and Recombinant DNA
Why? 1. Make large amounts of a
protein. 2. Study mutants 3. Improve
agriculture Yellow rice pest
resistance 4. Study and treat diseases.
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Cloning method to obtain a large amount of
a genetically identical population of DNA.
Recombinant DNA DNA from 2 sources.
Usually use a bacterium such as E. coli They
grow fast. This is what you put the recombinant
DNA in.
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Need to have a vector to get the DNA into the
bacterium. Virus (bacteriophage) Plasmid
(small piece of circular DNA in additon to
the main chromosome in bacteria.
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Plasmid small piece of circular DNA
Can insert a small piece of foreign DNA in a
plasmid using a restriction endonuclease
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Restriction endo nucleotides Nuclease cuts
DNA Endo in the middle Restriction specific
sequences
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EdoR1 hydrolyses DNA between G and A
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Sticky ends allow you to put it back together or
insert a new piece of DNA
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Plasmid small piece of circular DNA
Can insert a small piece of foreign DNA in a
plasmid using a restriction endonuclease
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Viruses
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Cut both phage and human DNA with a
restriction endonuclease
Cloning using a virus.
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Cloning with cells
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A DNA plasmid.
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How do we know that we have a cell that contains
a plasmid with our gene?
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3. What is genetic engineering?
The intentinal alteration of an organism at
the molecular level so that it exhibits altered
traits.
Useful in Agriculture Medicine Basic research
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Agriculture 1. Increased food production 2. Incr
eased shelf life 3. Drought resistance 4. Frost
resistance. 5. Increased milk production bovine
somatatrophin Look at Biochemical Connections
on p.343.
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• Problems?
• 1. Yellow rice.
• Seeds of genetically altered plants
• can not be reseeded.

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• Medical and health purposes.
• Genetically altered mosquitoes.
• Synthesis of human proteins (e.g. insulin)
• Treatment of diseases
• Cystic fibrosis
• Muscular dystrophy

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4. DNA libraries.
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How do you find the piece you want? Have a
piece of DNA that you know has the gene you want
go fishing with it.