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Extract DNA from an
organism’s cell to get the
GOI – gene of interest.
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Sequence the
gene.
Locate the gene.
Cut out the
gene using
restriction
enzymes.
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Remove plasmid from
bacterial cell.
Use restriction enzyme open
up the plasmid.
Use restriction enzyme on
the organism’s DNA (GOI)
to create sticky ends that are
complementary to the
plasmid.
Insert the GOI using ligase.
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What is the GOI?
What restriction enzyme is use to cut the DNA samples?
Why can the human and bacterium DNA combine?
What types of DNA are found in the bacterial cell?
What other genes may be found on the plasmid?
A Bacterial Plasmid:
What can you tell me
about the plamid?
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There are (6) restriction sites.
Find them.
There are (2) antibiotic
resistance genes. What
antibiotics?
The plasmid can replicate
when inside a bacterial cell that
is asexually reproducing
called binary fission.
Can insert a foreign gene.
Where? Which enzymes?
Bioluminescence and Fire flies
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Bioluminescence Surfing
The objective of this lab is to have you create a paper
recombinant plasmid. You will use colored paper,
scissors and tape. If you are successful, you will
have a two colored paper ring and pieces of colored
paper.
So what do you do with a recombinant plasmid?
Glowing Protein Transformation
Bozeman Transformation
Making Organisms Glow (Ted1)
What Other Organism Can Be Made to Glow (Ted2)
What Can be Done with Organisms (Ted3)
Cloning
Reason #1 – Make a transgenic organism.
Take out the copied genes in plasmid from the bacteria.
Take out the copied genes from the plasmids.
Put the gene in another organism’s genomic DNA
Reason #2 – Use to make a protein like a hormone.
Gene in the plasmid can be turned on by the bacteria or yeast cell
to make a protein.
Extract the protein and purify it for medical us.
In this lab, you will learn that a plasmid can have multiple
restriction sites to make a recombinant plasmid that can
contain antibiotic resistant genes and the gene of interest
(GOI). Your job is to cut the plasmid with specific enzymes
and determine the fragment size which is in base pairs (bp’s).
Objective of this lab is to
become familiar with the
different restriction
enzymes that can be
used to make a plasmid
vector
and
determine the fragment
size of a cut piece of
DNA
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Plasmid carries foreign DNA
into another organism.
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Virus carries foreign DNA
into a host cell of an
organism.
Vectors can be used to create Genetically Modified Organisms
Organisms that have acquired genes by artificial means .
If organism has two different species of DNA, it is a transgenic.
Vectors can be used to mass produce gene products (proteins)
Examples of proteins are insulin, human growth hormone,
blood clot dissolving enzymes, components for vaccines
Vectors can be used to in Gene Therapy (Virus as Vector only)
Gene Therapy is altering an afflicted person’s gene by
either replacing it or supplementing the defected gene
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