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Team:TU-Eindhoven/LEC/LabTheory
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<p>Since, the aim of this project is to design and produce a new multi-color display in which genetically engineered cells function as pixels analogous to how a flat panel display works and the decision has been made that yeast cells are, in this case, the most practical to work with, concerning multiple reasons discussed in section ‘Yeast versus E. Coli’, light emitting yeast cells which are sensitive to electric stimuli have to be engineered and produced. Considering this goal, the lab-team faces the challenge of introducing sensors sensitive to electrical stimuli resulting in an emission of light. After extensive literature research, it is hypothesized that CCH1-MID1 calcium plasma membrane channels, found in Saccharomyces cerevisiae but homologous to mammalian voltage-gated calcium channels, are able to facilitate a calcium influx upon plasma membrane depolarization<html><a href="#ref_Iida" name="text_Iida"><sup>[1]</sup></a></html>. Furthermore, it is known that GECO proteins<html><a href="#ref_zhao" name="text_zhao"><sup>[2]</sup></a></html> are sensitive to calcium resulting in fluorescence upon increased concentration. Regarding both, hypothesis and known fact, laboratory work could start.</p> | <p>Since, the aim of this project is to design and produce a new multi-color display in which genetically engineered cells function as pixels analogous to how a flat panel display works and the decision has been made that yeast cells are, in this case, the most practical to work with, concerning multiple reasons discussed in section ‘Yeast versus E. Coli’, light emitting yeast cells which are sensitive to electric stimuli have to be engineered and produced. Considering this goal, the lab-team faces the challenge of introducing sensors sensitive to electrical stimuli resulting in an emission of light. After extensive literature research, it is hypothesized that CCH1-MID1 calcium plasma membrane channels, found in Saccharomyces cerevisiae but homologous to mammalian voltage-gated calcium channels, are able to facilitate a calcium influx upon plasma membrane depolarization<html><a href="#ref_Iida" name="text_Iida"><sup>[1]</sup></a></html>. Furthermore, it is known that GECO proteins<html><a href="#ref_zhao" name="text_zhao"><sup>[2]</sup></a></html> are sensitive to calcium resulting in fluorescence upon increased concentration. Regarding both, hypothesis and known fact, laboratory work could start.</p> | ||
| + | <b>Nick: I'm working on this text, in Dropbox -> Lab approach.docx</b> | ||
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<p>DNA coding for CCH1, MID1 and the three colours (red, green, blue) GECO protein are obtained from H. Iida & K. Iida and Zhao, Campbell group via Addgene respectively. Via PCR these DNA strengths are ligated in vectors pBCT, YCpT and pYES3 respectively. Then, vectors coding for CCH1, MID1 and one of the three GECO colours were transformed into INVSc1 S. Cerevisiae cells using a heat shock protocol (more information can be found in section ‘Protocols’), obtaining three different strains of yeast, all containing the CCH1-MID1 calcium channel but each with a different colour GECO. </p> | <p>DNA coding for CCH1, MID1 and the three colours (red, green, blue) GECO protein are obtained from H. Iida & K. Iida and Zhao, Campbell group via Addgene respectively. Via PCR these DNA strengths are ligated in vectors pBCT, YCpT and pYES3 respectively. Then, vectors coding for CCH1, MID1 and one of the three GECO colours were transformed into INVSc1 S. Cerevisiae cells using a heat shock protocol (more information can be found in section ‘Protocols’), obtaining three different strains of yeast, all containing the CCH1-MID1 calcium channel but each with a different colour GECO. </p> | ||
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Revision as of 19:59, 25 September 2012
Overview
Since, the aim of this project is to design and produce a new multi-color display in which genetically engineered cells function as pixels analogous to how a flat panel display works and the decision has been made that yeast cells are, in this case, the most practical to work with, concerning multiple reasons discussed in section ‘Yeast versus E. Coli’, light emitting yeast cells which are sensitive to electric stimuli have to be engineered and produced. Considering this goal, the lab-team faces the challenge of introducing sensors sensitive to electrical stimuli resulting in an emission of light. After extensive literature research, it is hypothesized that CCH1-MID1 calcium plasma membrane channels, found in Saccharomyces cerevisiae but homologous to mammalian voltage-gated calcium channels, are able to facilitate a calcium influx upon plasma membrane depolarization[1]. Furthermore, it is known that GECO proteins[2] are sensitive to calcium resulting in fluorescence upon increased concentration. Regarding both, hypothesis and known fact, laboratory work could start.
Nick: I'm working on this text, in Dropbox -> Lab approach.docx
