Secrets of the rainforest

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Secrets of the Rainforest
A CANCER DRUG EXPLORATION
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
Tisha
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
ChromatographyHydrophobic 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 710
 Lesson 7- testing and placebos
 Lesson 8- Advocacy Groups and Viewpoints
 Lesson 9-Ethics and Economics
 Lesson 10- Preparing and Giving
Presentations
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