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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 Yizhi (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 and Molecular Biology teacher, Dr. Hackett. This year in Advanced Biotechnology and Molecular Biology, we began our year studying organic molecules and biofuels as an introduction to synthetic biology. We then moved on to protein structure and function, proceeding to learn different sequencing and genetic engineering techniques. Following this, we moved onto an epigenetics unit where we learned about the details of transcription and translation, including RNAi, as well as a deeper understanding of how our genes are regulated. In our next unit, we learned about the biology of cancer and the details of genome evolution. Finally, we learned about current advances in biotechnology and molecular biology including next-generation sequencing. In one of our projects this 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 techniques 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 induce the expression of fluorescent proteins. Throughout the year, we ran gels, performed restriction enzyme digests, sequenced DNA fragments, designed primers, ran PCR, and did genetic engineering. 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 had previously sequenced the GFP, YFP, mTangerine, and mCherry vectors as part of our course, so we knew the sequence for designing primers.

We designed primer sequences to amplify 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

Official Team Profile

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