Difference between revisions of "Part:BBa K1936001"
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===Usage=== | ===Usage=== | ||
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+ | While LOV2 domain was previously described for optogenetical control of gene expression (BBa_K1742000) and promotor characterization (BBa_K660004), 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. | ||
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+ | 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 (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] | ||
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+ | 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. | ||
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<html><dl class="figure"> | <html><dl class="figure"> | ||
<dt><img src="https://static.igem.org/mediawiki/2016/7/7c/Team_opoptosis_info_sheet_fig_4.png" alt="figure four" title="figure four" ></dt> | <dt><img src="https://static.igem.org/mediawiki/2016/7/7c/Team_opoptosis_info_sheet_fig_4.png" alt="figure four" title="figure four" ></dt> | ||
<dd><i>Figure 2: The LOV2-based optogenetic switch is activated by blue light.</i></dd> | <dd><i>Figure 2: The LOV2-based optogenetic switch is activated by blue light.</i></dd> | ||
</dl></html> | </dl></html> | ||
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===Biology=== | ===Biology=== |
Revision as of 22:06, 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 promotor characterization (BBa_K660004), 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 (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]
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 33
Illegal BamHI site found at 712
Illegal XhoI site found at 515 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1226