Team:Virtus-Parva Mexico/Parts
From 2014.igem.org
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HUMAN PRACTICES
"Science is not made for ouselves" (Niels Bohr in Copenhage)
For human practices we have done a variety of things, mostly dedicated to the dissemination of science within our University, locally in the city of Puebla and nationally through collaborations with other high schools
2>Law Initiative
First and foremost, we studied different procedures in order to synthesize the strongest and smallest magnetite particles we could make. Magnetite is a molecule that derives from iron, and has observable magnetic properties. Because we wanted to put together magnetite and DNA, we had to make them compatible, task we accomplished by functionalizing it with amino groups, that would allow it to form peptide bonds with our protein, HU. At the same time, our biology team was busy extracting, purifying and transforming E. Coli DNA to work with. This protein, HU, is a histone-like protein normally aids DNA into supercoiling around histones; the “Magnetic-Protein” complex we created mimics the nucleosome in DNA supercoiling process. This allows us to have a DNA “chromatin” with a magnetic core.
Combining magnetite, DNA and HU protein, we are building magnetic-core machines, which can be controlled through external electrical impulses. Because of the shape of our system, it is possible for it to have linear movement depending on the frequency applied to it, which has the potential to be incorporated into the medical sector as a pathogen-targeted therapy. This was our original idea and module one of our project.
As we were transforming our E. coli cells, we noticed it wasn’t as fast and efficient as we had hoped, which is how we came up with module two of the project. Quite simply, we wanted to take advantage of the shape of our system and its mobility thanks to magnetism in order to make a more efficient transformation. We were able to verify our method was more efficient by making cells express GFP and RFP, which can then be quantified with optic instruments.
The Idea
The basis for these “drills” will be NEMS, nano electro-mechanical systems, technology.
We took a survey to fellow iGEMers and external people in order to find out how many people knew about the existence of NEMS and if they knew how they worked. Turns out only 36% of survey takers had heard of the term before and of those, only 28% knew what it was!
Given these statistics, it became part of our project to teach newer generations about our subject.
NEMS
How exactly do NEMS come into play in our project?
Well, by combining an inorganically synthesized nanoparticle, called magnetite and DNA into what we call BioNEMS drill.
DNA Coiling into Chromosomes
The Making
Descripcion general de Seccion 3
Inorganic Section
After choosing the best method possible, it was time to silanize our magnetite in order for it to be biocompatible with DNA and be able to tie them together. In order for the silanization to take place, we used a solution of TEOS (tetraethoxysilane) dispersed in a medium of water and propanol and dripped this mix slowly onto our magnetite. Just like when we synthesized our particles, we tested different concentrations of TEOS and magnetite, as well as different addition rates in order to observe which combination would give us the smallest possible nanoparticles.
Our results were then characterized by DLS (dynamic light scattering), for which we observed a peak at 39 nm, once coated with TEOS, the peak was moved toward 60 and 80 nm. We also ran our two samples in the IR, comparing the spectra of the pure magnetite and silanized magnetite, we were able to distinguish a peak at 990.2 cm^-1 corresponding to a Si-O bond, confirming the correct silanization of the magnetite.