Difference between revisions of "Part:BBa K638401"
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Best used in an ''E. coli'' chassis such as strain [http://cgsc.biology.yale.edu/Strain.php?ID=111773 BW27783], with constitutive expression of an Arabinose transporter. | Best used in an ''E. coli'' chassis such as strain [http://cgsc.biology.yale.edu/Strain.php?ID=111773 BW27783], with constitutive expression of an Arabinose transporter. | ||
See our [[Part:BBa_I0500:Experience | experience of pBAD]] for some issues with tuning expression levels. | See our [[Part:BBa_I0500:Experience | experience of pBAD]] for some issues with tuning expression levels. | ||
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| + | We characterised this part using a GFP fusion - [[Part:BBa K638403]]. Unfortunately, Cambridge 2011 were unable to test export fully as of the wiki freeze (21st Sep). [http://2011.igem.org/Team:Cambridge/Experiments/Periplasmic_Export See our wiki] for more information on our experiments. | ||
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====Safety==== | ====Safety==== | ||
The protein coding sequence for reflectin originally came from cells of an edible squid. There have been no reported safety issues for reflectins, so we do not anticipate the need for extra precautions when using this BioBrick part. See our [http://2011.igem.org/Team:Cambridge/Safety safety page] for more information. | The protein coding sequence for reflectin originally came from cells of an edible squid. There have been no reported safety issues for reflectins, so we do not anticipate the need for extra precautions when using this BioBrick part. See our [http://2011.igem.org/Team:Cambridge/Safety safety page] for more information. | ||
Latest revision as of 23:59, 21 September 2011
Arabinose inducible TorA Export Tagged Reflectin A1 Poly His generator
This construct is a Reflectin A1 protein with a TorA signal peptide fused to the N terminus, which has been shown to allow export of GFP via the twin arginine pathway (see information on Part:BBa_K638402). In addition a poly his tag has been added to the C terminus and a pBAD promoter ( BBa_I0500) will control expression of the fusion protein on induction with arabinose. It is tightly controlled by two factors:
- L-arabinose monosaccharide taken up by the cell from the medium, which acts as an inducer.
- AraC protein included in the I0500 biobrick, which acts an a repressor.
Therefore, the araC-pBAD system offers regulatable control of gene expression in the presence of the inducer and highly repressed in the absence of the inducer. Read more about [http://2011.igem.org/Team:Cambridge/Experiments/Low_Level_Expression the pBAD and arabinose system in E.coli].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 1369 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
Usage and Biology
Best used in an E. coli chassis such as strain [http://cgsc.biology.yale.edu/Strain.php?ID=111773 BW27783], with constitutive expression of an Arabinose transporter. See our experience of pBAD for some issues with tuning expression levels.
We characterised this part using a GFP fusion - Part:BBa K638403. Unfortunately, Cambridge 2011 were unable to test export fully as of the wiki freeze (21st Sep). [http://2011.igem.org/Team:Cambridge/Experiments/Periplasmic_Export See our wiki] for more information on our experiments.
Safety
The protein coding sequence for reflectin originally came from cells of an edible squid. There have been no reported safety issues for reflectins, so we do not anticipate the need for extra precautions when using this BioBrick part. See our [http://2011.igem.org/Team:Cambridge/Safety safety page] for more information.