Team:TU-Eindhoven/LEC/LabTheory

From 2012.igem.org

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<p>Real-time imaging of biochemical events inside living cells is important for understanding the molecular basis of physiological processes and diseases<html><a href="#ref_merkx" name="text_merkx"><sup>[3]</sup></a></html>. Genetically encoded sensors based on fluorescent proteins (FPs) are frequently used for molecular recognition. In this iGEM project we use fluorescent proteins to provide the light in our display.</p>
<p>Real-time imaging of biochemical events inside living cells is important for understanding the molecular basis of physiological processes and diseases<html><a href="#ref_merkx" name="text_merkx"><sup>[3]</sup></a></html>. Genetically encoded sensors based on fluorescent proteins (FPs) are frequently used for molecular recognition. In this iGEM project we use fluorescent proteins to provide the light in our display.</p>
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[[File:Excitation spectra GECO.png|300px]] [[File:Emission spectra GECO.png|300px]]
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[[File:Making_GECO.png|600px]]
<p>A GECO is a protein which emits light in the presence of Ca<sup>2+</sup><html><a href="#ref_zhao" name="text_zhao"><sup>[2]</sup></a></html>. There are two important classes of genetically encoded Ca<sup>2+</sup> indicators. One is called the Forster Resonance Energy Transfer (FRET)-based cameleon type<html><a href="#ref_miyawaki" name="text_miyawaki"><sup>[4]</sup></a></html> and the other one is called the single Green Fluorescent Protein (GFP) type<html><a href="#ref_nakai" name="text_nakai"><sup>[5]</sup></a></html>. The GECO protein belongs to the single GFP type. Research has shown that Ca<sup>2+</sup> indicators targeted to the E. coli periplasm can be shifted toward the Ca<sup>2+</sup>-free or Ca<sup>2+</sup> -bound states by manipulation of the environmental Ca<sup>2+</sup> concentration<html><a href="#ref_zhao" name="text_zhao"><sup>[2]</sup></a></html>. Robert E. Campbell et al. named these Ca<sup>2+</sup> indicators GECOs. R-GECO, G-GECO and B-GECO emit red, green or blue light respectively, each with another  excitation and emission spectrum (Fig. 2 and Fig. 3).</p>
<p>A GECO is a protein which emits light in the presence of Ca<sup>2+</sup><html><a href="#ref_zhao" name="text_zhao"><sup>[2]</sup></a></html>. There are two important classes of genetically encoded Ca<sup>2+</sup> indicators. One is called the Forster Resonance Energy Transfer (FRET)-based cameleon type<html><a href="#ref_miyawaki" name="text_miyawaki"><sup>[4]</sup></a></html> and the other one is called the single Green Fluorescent Protein (GFP) type<html><a href="#ref_nakai" name="text_nakai"><sup>[5]</sup></a></html>. The GECO protein belongs to the single GFP type. Research has shown that Ca<sup>2+</sup> indicators targeted to the E. coli periplasm can be shifted toward the Ca<sup>2+</sup>-free or Ca<sup>2+</sup> -bound states by manipulation of the environmental Ca<sup>2+</sup> concentration<html><a href="#ref_zhao" name="text_zhao"><sup>[2]</sup></a></html>. Robert E. Campbell et al. named these Ca<sup>2+</sup> indicators GECOs. R-GECO, G-GECO and B-GECO emit red, green or blue light respectively, each with another  excitation and emission spectrum (Fig. 2 and Fig. 3).</p>
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[[File:Excitation spectra GECO.png|300px]] [[File:Emission spectra GECO.png|300px]]
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<h3>References</h3>
<h3>References</h3>

Revision as of 17:42, 26 September 2012