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From 2012hs.igem.org

Advisors

Jef Boeke and colleagues at Johns Hopkins University School of Medicine, including postdocs Leslie Mitchell and Patrick Cai, provided the design and inspiration for this project.

Jennifer Hackett (our teacher and former Johns Hopkins grad student) advised students as they worked on this project.

Course Overview

Our team is comprised of 5 juniors (Zoe M., Zoe E., Julia, Grace, and Kristen) from the Dalton school and our Advanced Biotechnology teacher, Dr. Hackett. This year in Advanced Biotechnology, we began our year studying organic molecules and biofuels as an introduction to synthetic biology. We then moved on to protein configuration and function, proceeding to learn different sequencing techniques. Following this, we moved onto an epigenetics unit where we learned about the details of transcription and translation as well as a deeper understanding of how our genes are inherited. In our final unit, we learned about the biology of cancer and the details of genome evolution. Throughout the course of the year, each student studied a specific iGEM project from a previous year and was asked to explain the ins and outs of the project. A major part of the course is revolved around actual lab work. We employed tecniques that we used in this iGEM project throughout the year in a variety of labs. For example, we tested to see if food was genetically modified and we modified protein vectors to include flourescent proteins. Throughout the year, we ran gels, performed restriction enzyme digests, sequenced DNA fragments to design primers, and ran PCR. All of these strategies were ultimately incorporated in our work on our portion of the iGEM project.

Attributions

Leslie Mitchell and Patrick Cai in Jef Boeke's lab at Johns Hopkins sent us the sequences of 30 yeast promoters to clone. In addition, we cloned the protein coding sequences of 6 fluorescent proteins that we had received as part of the BioBridge set distributed by Roger Tsien's lab (via Ann Sliski at Princeton University). These are BFP, YFP, GFP, mTangerine, mCherry, and mGrape1.

We designed primer sequences to clone each fragment, cloned them into pUC19, and confirmed their insertion into pUC19 by restriction digest.

Julia B. cloned:

promoters: MET15, GAL1, GAL1-L, GAL1-S, RAD52, RAD50

protein coding sequence: mTangerine

Zoe E. cloned:

promoters: RAD54, RAD55, MRE11, XRS2, REV1, TDH3

protein coding sequence: YFP

Grace F. cloned:

promoters: LIF1, MIG1, TUB1, CSE4, CLN2, CLB2

protein coding sequences: BFP, GFP

Kristen G. cloned:

promoters: ACT1, ENO1, ENO2, TEF2, TEF1, EAF6

protein coding sequence: mCherry

Zoe M. cloned:

promoters: HSP12, HSP26, HSC82, MSN2, MSN4, HHO1

protein coding sequence: mGrape1

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