1. Recombinant DNA Combining genes from two different species

2. Manipulating Genes Yesterday you found and translated jellyfish, coral and bacterial genes. Now what?

3. What are the steps for making a transgenic organism? What are the steps for making a transgenic organism?

5. How do we make the DNA? 1.Find the gene for the protein of interest, in both humans and goats. 2.Cut out the human gene. 3.Cut the goat gene to “open” it. 4.Insert the human gene into the opening.

6. Restriction Enzymes/Endonucleases Enzymes that cut DNA at specific sequences. “Sticky Ends” hhhh

7. How many pieces? Cut with: GAATTCCGGCCGAATTCAAGCTT CTTAAGGCCGGCTTAAGTTCGAA 1 1 2 2 3 3

8. How many pieces? Cut with: GAATTCCGGCCGAATTCAAGCTT CTTAAGGCCGGCTTAAGTTCGAA 1 1 2 2

9. What if your gene is the highlighted sequence? Cut with which? GAATTCCGGCCGAATTCAAGCTT CTTAAGGCCGGCTTAAGTTCGAA HindIII for sure, because EcoRI would cut your gene in two!

10. How many pieces? Cut with: GAATTCCGGCCGAATTCAAGCTT CTTAAGGCCGGCTTAAGTTCGAA 1 1 2 2 This is a “blunt cut” – harder to put back together again.

12. Vectors A “vector” is something that carries a specific gene into an organism. Plasmids are common for bacteria – and what we will use in our simulation.

13. Vectors Scientists have created special vector plasmids that have “polylinker” regions – lots of choice for cut sites!

14. Simulation Goal To take the _________________ genes for making proteins that __________________ in different colors (i.e. pink or green), and to insert them into _____________________ JELLYFISH or CORAL FLUORESCE BACTERIA !

15. Fluorescence Absorb energy at one wavelength (e.g. UV) and emit it at another (e.g. visible)

16. Normal Light UV Light

17. “Watching” the cell cycle: Cells are red during G1. Turn yellow at the start of replication. Turn green during S-phase

18. Studying Cancer with Fluorescence A mouse’s blood vessels were tagged with green. A human cancer tagged with red was injected into the mouse. Scientists can study how the cancer spreads, and how the blood vessels feed the cancer, to work toward a cure.

19. Tracking HIV’s virulence

20. Simulation: Cutting out Genes Materials: – DNA from yesterday, with highlighted genes – Scissors Your goal: cut your DNA in such a way that: – The jellyfish/coral gene has “sticky ends” on each side. – You “open” the bacterial plasmid. – The jellyfish/coral sticky ends “fit” in the open bacterial plasmid. (i.e. The bases are complementary.)

21. Clean Up Turn in the worksheet. Store DNA in Zip-loc for use tomorrow. (Include your names.)