Download IGEM BOOT CAMP

SUMMERCAMP2014
AIM
TO EXPRESS A
PLASMID VECTOR
IN A BACTERIA
AND DISPLAY
PHYSICAL
CHANGES
AIM
Understand recombinant DNA techniques, in particular the
transformation procedure.
Understand that DNA can be transferred to another organism and
therefore change the observable characteristics of that organism.
Become familiar with sterile technique and decontamination procedures
that are used to handle bacteria and also get familiarized with the
bioprocess involving upstream processing and downstream processing .
CONCEPT INVOLVED
Transformation of cells :The purpose of this technique is
to introduce a foreign plasmid into bacteria, the
bacteria then amplifies the plasmid, making large
quantities of it.
A plasmid is a small circular piece of DNA (about 2,000
to 10,000 base pairs) . Hence, the natural function of a
plasmid is to transfer genetic information vital to the
survival of the bacteria. It is this characteristic of
plasmids that is exploited for use in transformation.
CONCEPT INVOLVED
3:In molecular biology, transformation refers to a form of genetic exchange
in which the genetic material carried by an individual cell is altered by
incorporation of foreign (exogenous) DNA. This foreign DNA may be
derived from unrelated species and even other kingdoms, such as bacteria,
fungi, plants or animals, which would otherwise be inaccessible to an organism.
4:The bacterium we will be transforming, E.coli, lives in the human gut and
is a relatively simple and well understood organism. Its genetic material consists
mostly of one large circle of DNA 4-5 million base pairs (mbp) in length, with
small loops of DNA called plasmids, usually ranging from 5,000-10,000 base
pairs in length, present in the cytoplasm. It is these plasmids that bacteria can
transfer back and forth, allowing them to share genes among one another and
thus to naturally adapt to new environments.
DESIRED OUTPUT
The banana odor generator (BBa_J45200) catalyzes the conversion of
the precursor isoamyl alcohol to the odor isoamyl acetate that has a
banana smell.
Green fluorescent protein (GFP) from the bioluminescent
jellyfish Aequorea victoria When inserted into a plasmid and used
for the transformation procedure, the transformed bacteria will
express their newly acquired jellyfish gene and produce the
fluorescent protein, which causes them to glow green under
ultraviolet light.
LAB WORK
1:THE PLASMID/VECTOR WILL BE
FINALISED BY TODAY AND LAB WORK
WILL BE STARTED AS SOON AS POSSIBLE
2: INTRODUCTORY DETAILS ABOUT LAB
ETTIQUETES,EQUIPMENTS,PROCEDURES
HAVE BEEN DISCUSSED IN MEETINGS
PROGRESS
POINT
DISCUSSED
IN
MEETINGS
LAB WORK
•
•
•
•
INTRODUCTION TO LAB WORK
THEORY DETAILS
PLASMID/VECTOR TO BE INCOPORATED DISCUSSED
READINGS ABOUT UPSTREAM PROCESSING AND
DOWNSTREAM PROCESSING
• ESTIMATED TIME TO FINISH THE PROJECT IS WITHIN 2 WEEKS
• WILL BE STARTED AS SOON AS POSSIBLE