30-60 minutes 1.5 hour sessions are booked, but we may finish early

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6 Δεκ 2012 (πριν από 4 χρόνια και 7 μήνες)

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CURE 2009 Professional Development Workshops

Welcome to our professional
development workshop

CURE 2009 Professional Development Workshops

What is CURE?

Connecticut United for Research Excellence, Inc.



Represent biotechnology and pharmaceutical sectors before state
legislature and policy makers



Build a critical mass of biotech and pharmaceutical companies



Foster relationships between academic and industry research that lead
to technology transfer



Be the “go to” source for information about bioscience in Connecticut


CURE 2009 Professional Development Workshops

In Connecticut

Achillion Pharmaceuticals, Inc.

Alexion Pharmaceuticals, Inc.

Bayer Corporation

Boehringer Ingelheim Pharmeceuticals

Bristol
-
Myers Squibb

CGI Pharmaceuticals

CuraGen Corporation

HistoRx, Inc.

Institute for Pharmaceutical Discovery

Invitrogen Corporation

Marinus Pharmaceuticals, Inc.

Molecular Staging

Neurogen Corporation

Pfizer

Purdue Pharma L.P.

Rib
-
X Pharmaceuticals, Inc.

CURE 2009 Professional Development Workshops

Bioscience Careers

Connecticut Business and Industry Association

http://www.cbia.com/ed/STC/career_explorations/career_explor_info/health_bio.htm


“Biotechnology and
pharmaceutical companies are
among the fastest
-
growing
industries in Connecticut, yet they
are having difficulty finding
qualified workers with the right
technological and scientific skills
to fill high
-
demand, high
-
wage
jobs.”
-

CBIA

CURE 2009 Professional Development Workshops

Sponsors of BioBus Educational Programs

Platinum Sponsors
:

CURE and Connecticut Innovations

Gold Sponsors
:

Pfizer Inc and The Pfizer Foundation

Silver Sponsors
:

Boehringer Ingelheim

Bronze Sponsors
:

Achillion Pharmaceuticals, State of CT

Contributors
:

Bayer Corporation

Supporters
:

Wesleyan University, Quinnipiac


University, University of New Haven,


Yale University, SMART Technologies,


Bio
-
Rad, Invitrogen, Edvotek


CURE 2009 Professional Development Workshops



Generate student interest in and excitement for bioscience to
encourage career exploration




Provide educators with laboratory experience, bioscience
information and innovative teaching techniques




Generate public understanding, enthusiasm and support for
bioscience


CURE’s Educational Goals:

CURE 2009 Professional Development Workshops

CT BioBus Educational Programs

BioBus

BioConnection

Teacher Professional Development

Public Outreach/Events

CURE 2009 Professional Development Workshops




The BioBus has trained
over 800 teachers



Reached over 43,000
students



Conducted over 1,300
experiment sessions



As of 2008

CURE 2009 Professional Development Workshops

Today We Will:

Reinforce and increase your knowledge of bioscience



Help you to incorporate biotechnology in your curriculum


Provide hands
-
on bioscience opportunities for you and your
students

CURE 2009 Professional Development Workshops

LOGISTICAL INFORMATION FOR
YOUR BIOBUS VISIT

CURE 2009 Professional Development Workshops

Safety Protocol

ALL STUDENTS MUST:


Wear long pants and closed
-
toe shoes


Tie back long hair


Refrain from eating, drinking or gum chewing


Leave behind, jackets, hats, bags, purses or papers (everything
except BioBus notebook and pen/pencil)


Eat a meal and drink beforehand (breakfast is a must)

(Non
-
latex gloves and safety glasses

are provided)


CURE 2009 Professional Development Workshops

On board Teacher Responsibilities

The BioBus is an extension of YOUR classroom



Understand school emergency procedures and student
medical needs


Discipline


Pair up students
-

“standing” chart


Help instruct and demonstrate techniques


Be present on bus the ENTIRE session (have a plan/helper if
a student gets sick)


Make staff aware of any conditions/health issues you feel we
should know about (before visit day)



We are wheelchair accessible

CURE 2009 Professional Development Workshops

Things to Know…

Delivery and Parking Site:


The Bus is
usually

delivered the day/night before your visit
(occasionally the morning of)


Staff
cannot

move the bus once its parked!


The bus cannot be parked in fire zone


If wheelchair lift is used, the BioBus entrance must be 8 feet
from curb


Very level ground is needed for parking


2 spaces for staff vehicles near BioBus are appreciated, for
loading supply bins on to bus


CURE 2009 Professional Development Workshops

Things to Know…


Fill out class list form for every session


24 students MAX per
session



Break between sessions: 30
-
60 minutes



1.5 hour sessions are booked, but we may finish early (depends on
class size and skill level)



Make sure the pre
-
lab activities were done



Short tours can be arranged if requested four weeks beforehand


Check in with us on BioBus 30 minutes before first session


Staff arrives ~1 hour before first session, will not check in with school


CURE 2009 Professional Development Workshops

Press Releases

Samples available on our website

Press packets available on bus

Photo releases up to you

CURE 2009 Professional Development Workshops

Visiting Scientist Program

From a sponsor company


Scientific expert


Information on career opportunities


Demonstrate the applicability of the science


Resource for teachers and school


Parent scientists are welcome


Dr. Linda D. Strausbaugh, is a
Professor in the Dept. of Molecular
& Cell Biology at the University of
Connecticut

CURE 2009 Professional Development Workshops

After Your BioBus Visit


Sign up again!


Online request form


Training is good for 2 years


Consider BioConnection
Program




Thank You notes


“BioBus Sponsor”


Send to CURE offices


CURE 2009 Professional Development Workshops

Today: BioBus Experiments III



Glowing from Permutation



grades 9
-
12



1
-
2 day experiment





Genetically Modified Organisms



grades 9
-
12



2 day experiment


CURE 2009 Professional Development Workshops

Genetically Modified Organisms
(GMOs)


In this experiment, students will test soy
products to determine if they have been
genetically modified
. A company that grows
organic, non
-
GMO soybeans suspects that
some of their fields have been accidentally
planted with genetically modified Roundup
Ready
®

soybeans. Students will use a
scientific technique called
polymerase chain
reaction

(PCR) to amplify a specific portion
of the DNA to determine whether their
product does in fact contain GMOs.

CURE 2009 Professional Development Workshops

What is Roundup Ready
®
?



Plants containing the glyphosate resistance gene




Glyphosate is the active ingredient in the herbicide Roundup®



Inhibits an enzyme necessary for the synthesis of certain amino acids




Soy was the first Roundup Ready® crop produced by Monsanto




In 2004, approximately 85% of soy and 45% of corn grown in the U.S. were
grown from Roundup Ready® seed. (source: Greenomes.org)

CURE 2009 Professional Development Workshops

Genetically Modified Organisms

Organisms that are modified by the alteration
or introduction of foreign DNA, in order to
produce a desired trait or effect


Plants are the most common targets
(transgenic crops):


Pest resistance (e.g. Bt
-
hybrid corn)


Drought resistance


Cold
-
tolerance


Shelf
-
life


Ripening


Nutritional value




Bt
-
hybrid vs. non
-
Bt
-
hybrid corn

CURE 2009 Professional Development Workshops

Creation of a GMO

Genetic material is artificially combined, usually in
the form of circular pieces of DNA called
plasmids


Plasmids contain not only DNA coding for the
trait of interest, but also DNA that controls the
expression and selection of the trait


Delivery methods:


Agrobacterium tumefaciens


Ballistics or gene gun


Electroporation


Chemical and/or heat treatment




http://sustain.no/virtue/newsletter/00_08/curr
-
trant/more
-
info/plasmid.jpg

CURE 2009 Professional Development Workshops

Restriction enzymes



Isolated from bacteria




Microscopic scalpels




Cut DNA molecules at specific
sequences known as “recognition
sites”

Eco R1: from
E. coli
,
strain R1


Cuts the

Following:


…G A A T T C…

…C T T A A G…

…G

A A T T C…

….C T T A A

G…

CURE 2009 Professional Development Workshops


Process by which genetic material carried by an individual cell
is altered by the addition and expression of foreign DNA


The DNA may or may not be incorporated into the genome



Electrical, thermal, mechanical and chemical and
biological

methods




Transformed cells are identified by some type of marker or trait


Antibiotic resistance


Color

Transformation

CURE 2009 Professional Development Workshops

Transformation Using
Agrobacterium
tumefaciens



Bacterial species



Normally infects plants and
causes crown
-
gall disease



Very efficient at inserting DNA
into plant cell genome


Bingo!


CURE 2009 Professional Development Workshops

T
-
DNA leads to synthesis of:


Auxin


plant hormone causes cell proliferation


Opines


serve as a source of energy for
A. tumefaciens

Symbiotic Relationship

CURE 2009 Professional Development Workshops

Step 1. Engineer
Agrobacterium

with your
favorite gene(s)….

Ti = Tumor inducing

Step 2. Disarm the genes that cause crown gall
disease

CURE 2009 Professional Development Workshops

http://dragon.zoo.utoronto.ca/~jlm
-
gmf/T0301C/images/transgenicflowchart.jpg

Step 3: Infect plant cells with engineered
Agrobacterium…

CURE 2009 Professional Development Workshops

Recombinant DNA Animation

CURE 2009 Professional Development Workshops

PBS Harvest of Fear

CURE 2009 Professional Development Workshops

Detecting a GMO

ELISA
-

Uses antibodies to detect the
presence of foreign proteins



Processing can sometimes destroy the
protein


PCR

-

detects and amplifies foreign DNA



More sensitive than ELISA


Can be used on processed foods


More general: usually detects common
plasmid element such as 35S promoter



Photo credit: Jon Morris

GMO

Promoter
: a regulatory region of DNA located upstream (towards the 5’ region) of a gene, providing a
control point for regulated gene expression.

CURE 2009 Professional Development Workshops

PCR


P
olymerase
C
hain
R
eaction



Goal: Amplify a target sequence of
DNA



Mimics the natural process of DNA
replication



DNA is replicated in minutes by
repeated heating and cooling of
reaction tubes

CURE 2009 Professional Development Workshops

PCR Components


DNA template


Two Primers (segments of DNA that define the
region to be copied)


Taq polymerase (from the bacteria
Thermus
aquaticus)


Free nucleotides (dNTPs) A T G C


Buffer containing Mg
2+


Thermal cycler



CURE 2009 Professional Development Workshops

DNALC PCR Animation

CURE 2009 Professional Development Workshops

Bio
-
Rad PCR Animation

CURE 2009 Professional Development Workshops

DNA Interactive

CURE 2009 Professional Development Workshops

Gel Electrophoresis


What does it mean?


Gel

-

scientific Jell
-
O

Electro

-

electricity

Phoresis



to carry across

Gel electrophoresis uses electricity to carry molecules
across a gel


CURE 2009 Professional Development Workshops

Genetic Science Learning Center


CURE 2009 Professional Development Workshops

Pre
-

and Post
-

lab activities

Pre
-
Lab Activities


Bioinformatics: Electronic PCR


DNA fruit extraction


DNA transcription and translation exercise


PCR animations and related websites


Post
-
Lab Activity



GMO


CAPT Generation III Embedded Task



CURE 2009 Professional Development Workshops

BLAST

CURE 2009 Professional Development Workshops

GMO PCR Primers

Forward primer




5’
-


GCT CCT ACA AAT GCC ATC A
-

3’


Reverse primer


5’
-


GAT AGT GGG ATT GTG CGT CA

-

3’

CURE 2009 Professional Development Workshops

35S Promoter Sequence

Forward

Primer

Reverse

Primer

gctcctacaa

atgccatca
t tgcgataaag gaaaggccat cgttgaagat gcctctgccg
acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagacgttc
caaccacgtc ttcaaagcaa gtggattgat gtgatatctc cactgacgta aggga
tgacg

cacaatccca ctatc

Amplicon: 195 base pairs

CURE 2009 Professional Development Workshops

On to the BioBus!

CURE 2009 Professional Development Workshops

Glowing From Permutation

What is genetic engineering? How is it
done? In this experiment students insert
the gene for Green Fluorescent Protein
(GFP) into
E.coli

bacteria using a scientific
technique called
transformation
. The
next day, students will be able to see if
they successfully created transformed
bacteria, as these clones will fluoresce
green! This experiment highlights the
importance of experimental controls and
the processes of DNA transcription and
translation.

BioRAD

CURE 2009 Professional Development Workshops

Genetic Engineering


Manipulation of DNA



The DNA of interest is often “inserted” into a plasmid.



The plasmid is inserted into a host cell (transformation)



Replication of the cell = replication of plasmid



Bacteria produce lots of your DNA (and protein) of
interest





CURE 2009 Professional Development Workshops

A typical bacterial cell

Picture taken from: http://www.biosci.uga.edu/almanac/bio_103/notes/may_30.html.

Graphic from
www.accessexcellence.org

Cloning into a Plasmid

CURE 2009 Professional Development Workshops

Bacterial Transformation Animations

CURE 2009 Professional Development Workshops

DNA Interactive

CURE 2009 Professional Development Workshops

Pre
-

and Post
-

Lab Activities

Pre
-
Lab Activities
-



Introduction to Restriction Enzymes


Restriction Enzymes and Bacterial Transformation


Cloning GFP into a plasmid


Can I genetically transform an organism?


Practice streaking a plate

After the BioBus
-



Review Questions


Data Collection and Analysis


Analysis Questions

Post
-
Lab Activities


Investigation into Diabetes Mellitus


Molecular Toolbox


CAPT Embedded Task

CURE 2009 Professional Development Workshops

Back to the BioBus!

CURE 2009 Professional Development Workshops

Questions?

Thank you!