


Uses a series of solutions that lyse the cell and a hot water bath to destroy
nucleases followed by using 95% cold ethanol to precipitate the DNA.
Extracted DNA contains organism’s cell to get the GOI – gene of interest.
GOI removed from the genomic DNA and inserted into another type of DNA.



Identify the GOI by determining
its nucleotide sequence
Select a certain restriction
enzyme that can cut around the
gene. The restriction site
should not be in the gene. If
restriction enzyme cuts in gene,
gene may not function.
Perform a restriction digest to
cut out the GOI.




Remove plasmid from
bacterial cell.
Use restriction enzyme
open up the plasmid and
create sticky ends.
Use restriction enzyme on
the organism’s DNA (GOI)
to create sticky ends that
are complementary to the
plasmid sticky ends
Insert the GOI into the
plasmid by using ligase.
1.
2.
3.
4.
5.
A Bacterial Plasmid’s Parts
What part makes the bacteria antibiotic resistant?
What gene is needed for it to replicate?
How many restriction sites are present?
Where don’t you want to cut ever?
If the plasmid were digested with SaII, how many fragments would you get?
1.
2.
3.
4.
5.
6.
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?
Find the recombinant plasmid in the diagram.
Bioluminescence and Fire flies
Top Ten Bioluminescent Organisms
Bioluminescence Surfing
The objective of the Paper Plasmid lab is to have you create a paper
recombinant plasmid, a plasmid with a new gene inserted. The
plasmid will contain DNA from two different organisms. You will use
colored paper, scissors and tape to do this. If you are successful,
you will have a two colored paper ring and extra pieces of paper.

Put it back into a bacterial cell
Now the new cell has a new trait because the plasmid has
the new gene in it. It has a new phenotype.
In the above illustration, find the bacteria with a plasmid and find
The bacteria that do not have a plasmid. The one with the plasmid
can be used to clone the inserted gene.
What is Gene Cloning?
Above is a picture of millions of bacterial cells. They all
Are glowing which means each cell has taken up the
plasmid with the lux gene. What would a nonglowing
colony of cells tell you about the bacteria?
Watch the Transformation Procedure
DNA Cloning
Glowing Protein Transformation
bozeman Transformation
Learn how insulin is made using recombinant plasmid
Here is another illustration of how to clone a gene using a recombinant plasmid….