Difference between revisions of "Part:BBa K1936001"
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While LOV2 domain was previously described for optogenetical control of gene expression (<html><a href="https://parts.igem.org/Part:BBa_K1742000">BBa_K1742000</a></html>) and as reporter gene (<html><a href=" https://parts.igem.org/Part:BBa_K660003">BBa_K660003</a></html>), we took a new approach. The described fusion protein Tom5-eGFP-LOV2 is designed to recruit proteins of choice to the outer mitochondrial membrane (OMM) in engineered eukaryotic cells. | While LOV2 domain was previously described for optogenetical control of gene expression (<html><a href="https://parts.igem.org/Part:BBa_K1742000">BBa_K1742000</a></html>) and as reporter gene (<html><a href=" https://parts.igem.org/Part:BBa_K660003">BBa_K660003</a></html>), we took a new approach. The described fusion protein Tom5-eGFP-LOV2 is designed to recruit proteins of choice to the outer mitochondrial membrane (OMM) in engineered eukaryotic cells. | ||
− | Tom5 is a part of the eukaryotic translocase of outer membrane protein complex and has a single membrane anchor unit that integrates into OMM. [1] We fused it to the N-terminal end of LOV2 domain from <html><i>A.sativa</i></html>. Upon exposure to blue light (<html>λ</html>=473nm) the LOV2 peptide undergoes a conformational change and exposes its C-terminal J-alpha-helix which is then able to bind a specifically engineered PDZ domain (<html><a href="https://parts.igem.org/Part:BBa_K1470005">BBa_K1470005</a></html>). [2] | + | Tom5 is a part of the eukaryotic translocase of outer membrane protein complex and has a single membrane anchor unit that integrates into OMM. [1] We fused it to the N-terminal end of LOV2 domain from <html><i>A.sativa</i></html>. Upon exposure to blue light (<html>λ</html>=473nm) the LOV2 peptide undergoes a conformational change and exposes its C-terminal J-alpha-helix which is then able to bind a specifically engineered PDZ domain (<html><a href="https://parts.igem.org/Part:BBa_K1470005">BBa_K1470005</a></html>). [2] The fused enhanced green fluorenscence protein (eGFP) allows the tracking of cellular localization by microscopy. |
− | + | ||
− | The fused enhanced green fluorenscence protein (eGFP) | + | |
Our blue light dependent system can be used to recruit any protein of interest to the outer mitochondrial membrane by simply fusing it to an ePDZ domain. | Our blue light dependent system can be used to recruit any protein of interest to the outer mitochondrial membrane by simply fusing it to an ePDZ domain. |
Revision as of 23:10, 15 October 2016
Tom5-eGFP-LOV2
ALERT !!!Under construction !!
Usage
While LOV2 domain was previously described for optogenetical control of gene expression (BBa_K1742000) and as reporter gene (BBa_K660003), we took a new approach. The described fusion protein Tom5-eGFP-LOV2 is designed to recruit proteins of choice to the outer mitochondrial membrane (OMM) in engineered eukaryotic cells.
Tom5 is a part of the eukaryotic translocase of outer membrane protein complex and has a single membrane anchor unit that integrates into OMM. [1] We fused it to the N-terminal end of LOV2 domain from A.sativa. Upon exposure to blue light (λ=473nm) the LOV2 peptide undergoes a conformational change and exposes its C-terminal J-alpha-helix which is then able to bind a specifically engineered PDZ domain (BBa_K1470005). [2] The fused enhanced green fluorenscence protein (eGFP) allows the tracking of cellular localization by microscopy.
Our blue light dependent system can be used to recruit any protein of interest to the outer mitochondrial membrane by simply fusing it to an ePDZ domain.
- Figure 2: The LOV2-based optogenetic switch is activated by blue light.
Biology
Characterization
References
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1226