Extract DNA from an organism’s cell to get the GOI – gene of interest.

Sequence the gene.

Locate the gene.

Cut out the gene using restriction enzymes.

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.

1. What is the GOI?2. What restriction enzyme is use to cut the DNA samples?3. Why can the human and bacterium DNA combine?4. What types of DNA are found in the bacterial cell?5. What other genes may be found on the plasmid?

1.There are (6) restriction sites. Find them.2.There are (2) antibiotic resistance genes. What antibiotics?3.The plasmid can replicate when inside a bacterial cell that is asexually reproducing called binary fission.4.Can insert a foreign gene. Where? Which enzymes?

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.

Bioluminescence and Fire flies

Top Ten Bioluminescent Organisms

Bioluminescence Surfing

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 cellto 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

anddetermine the fragment size of a cut piece of DNA

Plasmid carries foreign DNA into another organism.

Virus carries foreign DNA into a host cell of an organism.

Vectors can be used to create Genetically Modified OrganismsOrganisms 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 byeither replacing it or supplementing the defected gene

GMO Rice and Vitamin A Deficiency

Brief on GMO Rice

Ti Plasmid (MacGraw Hill)

Using Plasmids to Make Insulin