Secrets of the Rainforest

The Mysterious Green Florescent Leaves

While Hiking through the Andean Rain Forest, You and a friend, Tisha, obtain medicinal green leaves from Ramon

You bring back to Biotex, a biotech company working on a cure for cancer

Hi, I am Tish

a

Green Florescent Protein

Biotex scientists determine that the leaves contain a florescent green protein

DNA from the leaves is inserted into bacterial cells

Cloning the Gene from Mysterious Leaves

More on Cloning the Gene…

Purifying the GFP

Biotex technicians need to separate the GFP from other bacterial proteins

Column Chromatography separates contaminating proteins from GFP

The result is a purified GFP that can be tested for its ability to cure stomach cancer

Testing and Placebos

GFP will need to be tested on rats before it is allowed for a human trial

The US Food and Drug Administration will examine Biotex’s experiments to determine if the protein is safe and cures stomach cancer without negative side effects

Ethics and Economics

Once GFP is approved by the FDA, you will develop GFP as a commercial product

You will need to be aware of the following viewpoints: FDA, social conscience, patient advocacy groups, and the employees of Biotex

You must resolve the dilemmas in order to successfully market GFP

Final Presentations

Turn in all lab work with analysis questions

Video Presentation of your marketing of GFP Address issues of all

dilemmas 1-8

Background Information- Lesson 1

How to use Pipettes

Streaking of agar plates to Produce Single Colonies (Sterile Technique)

Background Continued…Lessons 2 & 3

Process of Transformation (GMOs or genetic recombinants)

Jellyfish , GFP and pGLO and Fluorescence

Isolating Clones to Propagate in Liquid Culture Media

Background Continued…Lesson 4

Concentration of Bacteria from Liquid Culture Media

Background Continued…Lesson 5

Prepare a Bacterial Lysate for Protein Chromatography

Background Continued…Lesson 6

Protein Chromatography-Hydrophobic Interaction Chromatography (HIC)

GFP is hydrophobic and will stick to hydrophobic beads in salt water buffer

When salt is removed, GFP will then be collected

4 buffers: 1) Equilibration

buffer (salt buffer) 2) Binding Buffer

(high salt) Wash buffer (medium

salt) Elution buffer (low

salt)

Background Continued…Lessons 7-10

Lesson 7- testing and placebos Lesson 8- Advocacy Groups and

Viewpoints Lesson 9-Ethics and Economics Lesson 10- Preparing and Giving

Presentations