Biotech Lab #1 -Extraction

1. Biotech Lab #1 -Extraction • Extract DNA from an organism’s cell to get the GOI – gene of interest.

2. Biotech Lab#2 – Restriction Enzymes • Sequence the gene. • Locate the gene. • Cut out the gene using restriction enzymes.

3. Biotech Lab #3 – Recombinant Plasmid • 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.

4. 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?

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

6. The Lux (Luciferase) Gene Bioluminescence and Fire flies Top Ten Bioluminescent Organisms 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?

7. Transforming Bacteria Glowing Protein Transformation Bozeman Transformation

8. Can a glowing gene be placed in a eukaryote?Click on this link: Glowing Sheep in the News

9. What’s being done in the world of biotechnology? Making Organisms Glow (Ted1) What Other Organism Can Be Made to Glow (Ted2) What Can be Done with Organisms (Ted3)

10. Why make a recombinant plasmidandtransform bacteria? Cloning

11. 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. What is done with cloned genes?What is done with recombinant plasmid that is used in a transformation?

12. Biotech lab #4: Making the vectorClick on this link to see how it’s doneConstructing a Plasmid Vector 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).

13. DNA Scissors Part II: Digesting a Plasmid 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

14. Vector is something that is used to carry something and deposit it in that place • 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 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

15. Using a plasmid as a vector: Ti Plasmid (MacGraw Hill) Brief on GMO Rice GMO Rice and Vitamin A Deficiency Using Plasmids to Make Insulin