Team:TU-Munich/HumanPractice/Education

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Revision as of 08:53, 3 October 2013

Introducing Synthetic Biology Kits at educational institutions

Introduction

Prepared School-Kits, ready for delivery!

Corresponding to this famous statement in pedagogics, the TU Munich iGEM Team of 2013 realized the importance to inspire and teach disciples in educational institutions to long for the endless immensity of Synthetic Biology. We want to make it possible for disciples to take their first steps in the enormously evolving field of Synthetic Biology and let them take a breath of this subject, which will change the world in the near future due to its unlimited fields of application.

The idea is to send an "Educational-Kit" with all required reagents and lab-protocols, based on the experiments we designed, to educational institutions which are interested in inspiring young prospective people to think about a promising career in science. Our experiments are styled constitutive, based on every previously performed experiment, so every disciple, independent of training level, is able to understand and take notice of the resulting phenomena which Synthetic Biology makes possible.

The Educational-Kit

The reagents, which are necessary for the realization of the experiments are all found in the kit, except for competent cells, which have to be stored at -20°C. The kit includes the following reagents:

Reagent	Amount of the Reagent	Is it existent in the kit?
LB-Medium	Reagents for about 3 L are in the kit	Yes
"Banana-Odor"-Plasmid	100 µl (10 ng/µl)	Yes
"RFP-Generator"-Plasmid	100 µl (10 ng/µl)	Yes
"Luciferase"-Plasmid	100 µl (10 ng/µl)	Yes
Isoamylalcohol (>98 %)	10 ml	Yes
Agar plates (Resistance: Kanamycin)	4 Plates	Yes
Agar plates (Resistance: Chloramphenicol)	2	Yes
L-Arabinose (500 mM)	3.76 g (50 ml)	Yes
Kanamycin	25 ml	Yes
Chloramphenicol	25 ml	Yes
Competent Cells	6 Tubes (Aliquotvolume: 150 µl)	No

As you may have noticed, we provide three different plasmids, which are used seperately in each designed experiment. The aim of the experiments is to perform the whole way from transformation, over cultivating cells, inducing the production of proteins and in the end demonstrate the conversion of a substrate by the produced enzyme.

Experiment Number 1

Results of Experiment Number 1

The aim of the first experiment is to transform the "RFP-Generator"-Plasmid into Escherichia coli containing genes coding for Chloramphenicol resistance and a red fluorophor and to plate them on Chloramphenicol-Agarplates. The Agarplates, provided with the antibiotica Chloramphenicol, make a selection process possible, whose aim is to kill all bacteria which have not incorporated the "RFP-Generator"-Plasmid. The expression rate of the red fluorophor is controlled by a constitutive promotor, so the Red fluorescent Protein is expressed continuously. After just about one day of incubation time, the disciples are able to notice a change in colour of the bacteria from ocher to red.

Experiment Number 2

Structure of the Luciferase

In the second experiment disciples should perform the transformation, learned in experiment one, with the "Luciferase"-Plasmid which encodes genes for Kanamycin resistance and a protein-fluorophor called Luciferase. Another very important point is that the Luciferasegene is subordinated to a L-Arabinose Operon. After the transformation of the DNA-Plasmid has taken place, discicples should learn how to cultivate bacteria in liquid LB-Medium and how to induce protein expression by using L-Arabinose as an inductor of the L-Arabinose promotor. The glow of the Luciferase can be observed after just a couple of hours.

Experiment Number 3

Results of Experiment three

In the third experiment, all aspects learned in the previous experiments come together. They are necessary to understand the following approaches. The "Banana-Odor"-Plasmid encodes an enzyme called Alcohol-Acetyltransferase I. This enzyme is able to convers Isoamylalcohol into Isoamylacetat, which spreads the odor of banana. Additionally there is found a resistance gene for Kanamycin and an inducible L-Arabinose promotor, which controls the expressions rate of the Alcohol-Acetyltransferase I. The disciples should transform the "Banana-Odor"-Plasmid into Escherichia coli, cultivate the successfully transformed bacteria in a bigger scale and in the end, induce the protein expression using L-Arabinose. In the last step, the disciples should add the substrate Isoamylalcohol to the cell culture, so the produced enzyme can transform it into Isoamylacetat.

Target Audience

Our target audience have been secondary schools, which focus a biotechnical educational pathway, so all students have access to the required laboratory equipment for the experiments. Before sending the kits to the schools, we tested them in detail - with success. All schools we contacted, to spread our Synthetic Biology kits, are very enthusiastic concerning our idea of supporting young prospective scientists, so they are very excited to receive and use our kit!

Availability of the School-kit

We appreciate other interested schools in trying our “School kits”. Do not be ashamed of contacting us, so we can supply you with our forward-looking Synthetic Biology “School-kit”!

References

http://www.ncbi.nlm.nih.gov/pubmed/6327079 Edens et al., 1984

http://www.ncbi.nlm.nih.gov/pubmed/6327079 Edens et al., 1984 Edens, L., Bom, I., Ledeboer, A. M., Maat, J., Toonen, M. Y., Visser, C., and Verrips, C. T. (1984). Synthesis and processing of the plant protein thaumatin in yeast. Cell, 37(2):629–33.

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-===Experiment Number 1===
+==Experiment Number 1==
 [[File:TUM13_RFP_Generator.jpg|thumb|right|400px|Results of Experiment Number 1]]
 The aim of the first experiment is to transform the "RFP-Generator"-Plasmid into ''Escherichia coli'' containing genes coding for Chloramphenicol resistance and a red fluorophor and to plate them on Chloramphenicol-Agarplates. The Agarplates, provided with the antibiotica Chloramphenicol, make a selection process possible, whose aim is to kill all bacteria which have not incorporated the "RFP-Generator"-Plasmid. The expression rate of the red fluorophor is controlled by a constitutive promotor, so the Red fluorescent Protein is expressed continuously. After just about one day of incubation time, the disciples are able to notice a change in colour of the bacteria from ocher to red.