Transcript A Bacterial Plasmid: What can you tell me about the plamid?

Biotech Lab #1 -Extraction
• Extract DNA from an
organism’s cell to get the
GOI – gene of interest.
Biotech Lab#2 – Restriction Enzymes
• Send out DNA to
determine the gene
sequence
• Locate the gene.
• Cut out the gene
using restriction
enzymes.
Biotech Lab #3 – Recombinant Plasmid
• Remove plasmid from bacterial
cell.
• Use restriction enzyme (RE)
open up the plasmid.
• Use restriction enzyme to cut
the gene out of on the
organism’s DNA. Create sticky
ends that are complementary
to the plasmid’s sticky ends.
• Insert the gene using ligase.
How does one determine which
RE’s to use?
Illustration of Making a Recombinant Plasmid
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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?
The Luciferase Gene
Bioluminescence and Fire flies
Top Ten Bioluminescent Organisms
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.
A Bacterial Plasmid:
What can you tell me
about the plamid?
How many restriction sites?
How many antibiotic resistant
genes?
Can this plasmid make more copies
of itself? Why? When?
Is this a recombinant plasmid?
Explain.