dna technology - 2. what are plasmids? small, circular dna molecules they are separate from the...
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DNA Technology - 2
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What are plasmids?
Small, circular DNA molecules
They are separate from the bacterial chromosome
(Found in bacteria)
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Why are plasmids useful?
They are small
And, are easily taken up by bacterial cells
When they are taken up they are called vectors
vector A DNA carrier that move genes from one cell to another
They are used to manipulate genes in the lab
What happens when the bacterial cell replicates its chromosome?
It also replicates the plasmid DNA
Contain genes useful to the bacteria
(including any foreign DNA as well)
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What does cutting and pasting of DNA mean?
Enzymes are involved
For cutting, what are they called?
Restriction enzymes
For pasting, what are they called?
DNA ligases
They recognize specific sequence of: A T C G
e.g., A T C G
Hundreds have been isolated from bacteria
They bind cut ends back together
(by covalent bonds between adjacent nucleotides)
Its the last step to make recombinant DNA
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So, this cutting and pasting of DNA is used to:
Cut gene sequence of interest (e.g., from human DNA)
Cut a plasmid (from a bacterium)
Create a recombinant DNA molecule
By using the plasmid
And ‘pasting’ a human gene sequence into it
Enzyme name?
Enzyme name?
Enzyme name?
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Polymerase Chain Reaction
Or PCR
• In which any segment of DNA can be copied
• Quickly and precisely
• PCR is a technique
Why is it so important?
Use minute amounts of blood or other tissue
To generate enough DNA for analysis
e.g., DNA from the follicle of ONE stand of hair
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How does PCR work?
Make a mixture of:
• the DNA sample, • some nucleotides, • an enzyme, DNA polymerase
Treat the mixture to:
• cycles of heating Allows separation of DNA strands
• cycles of coolingAllows DNA strands to re-form duplexes
DNA Replication occurs during cooling cycle
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Within a few hours:
In each cycle, the DNA is DOUBLED
___1st cycle
___2nd cycle
___3rd cycle___4th cycle
How many copies of DNA after each cycle?
How many copies after a 5th cycle?
PCR can generate billions of copiesFrom a SINGLE DNA molecule
Enough to do extensive analyses
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The Human Genome Project
What was the goal of HGP?To determine the nucleotide sequence all the DNA
In any given human cell
To identify the location & sequence of every gene
1990 - 2003
What was discovered?Our DNA contains ~ 2.9 billion nucleotide pairs
About 25,000 genes
There is a LOT of DNA that isn’t made up of genes
About 97% is non-coding DNA
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Learning check
1. Why is only the slightest trace of DNA at a crime scene often sufficient for forensic analysis?
2. A carrier that moves DNA from one cell to another, such as a plasmid, is called a ________
3. What features of a DNA fragment causes it to move through a gel during electrophoresis?
a. Its nucleotide sequenceb. The hydrogen bonds between its base pairsc. Its double helix shape d. The electrical charges of its phosphate groups
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4. A paleontologist has recovered a bit of organic material from the 400 year old preserved skin of an extinct dodo. She would like to compare DNA from the sample with DNA from living birds. The most useful method for increasing the amount of dodo DNA available for testing is __________
5. Why is golden rice pale yellow in color?
a It is rich in chlorophyll a.
b It is nutrient-poor.
c It is rich in beta-carotene.
d It is rich in chlorophyll b.
e It is rich in phycobilins.
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What does each ‘band’ consist of?
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What does this figure show?
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Some Review
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Structure and Function?
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1. Name molecule2. Name molecule3. Name molecule4. Name the reaction
5. Name of molecule6. What does the arrow refer to?7. Name of molecule8. Name of molecule9. Where does this take place?
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