Difference between revisions of "Part:BBa K2387053"

 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K2387053 parameters</partinfo>
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In order to test the functionality of this split protein under the araC/pBad promoter, the absorbance and the fluorescence spectra of the full length protein and the split protein were measured after induction with 0.002% arabinose (Figure 1). The stability of the shape in both spectra for the full length protein and the split protein show that the split has been successful.   
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[[file:T--Wageningen_UR--Results_Spectra.jpg|800px|center|thumb|<p align="justify">'''Figure 1: Absorbance and fluorescence spectra of full length mCerulean (left) and split mCerulean (right), measured after induction with 0.002% arabinose.'''</p>]]
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This protein is part of a collection of split proteins developed for BiFC analysis and tested with the CpxR system ([https://parts.igem.org/Part:BBa_K2387032 BBa_K2387032]). These proteins were characterized through multiple experiments using as interacting proteins antiparallel synthetic leucine zippers. The proteins of this collection are the ones found in Table 1.
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<table style="width:100%">
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<tr>
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<th colspan="3">Table 1: Split Proteins.</th>
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</tr>
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<tr>
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<th>Protein</th>
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<th>Part Number (Full length)</th>
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<th>Part Number (Split Protein)</th>
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</tr>
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<tr>
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<td>mRFP</td>
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<td>[https://parts.igem.org/Part:BBa_K2387054 BBa_K2387054]</td>
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<td>[https://parts.igem.org/Part:BBa_K2387055 BBa_K2387055]</td>
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</tr>
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<tr>
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<td>eYFP</td>
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<td>[https://parts.igem.org/Part:BBa_K2387003 BBa_K2387003]</td>
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<td>[https://parts.igem.org/Part:BBa_K2387065 BBa_K2387065]</td>
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</tr>
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<tr>
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<td>mVenus</td>
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<td>[https://parts.igem.org/Part:BBa_K2387045 BBa_K2387045]</td>
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<td>[https://parts.igem.org/Part:BBa_K2387046 BBa_K2387046]</td>
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</tr>
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<tr>
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<td>sfGFP</td>
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<td>[https://parts.igem.org/Part:BBa_K2387047 BBa_K2387047]</td>
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<td>[https://parts.igem.org/Part:BBa_K2387048 BBa_K2387048]</td>
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</tr>
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<tr>
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<td>mCerulean</td>
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<td>[https://parts.igem.org/Part:BBa_K2387052 BBa_K2387052]</td>
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<td>[https://parts.igem.org/Part:BBa_K2387053 BBa_K2387053]</td>
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</tr>
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</table>

Revision as of 11:48, 31 October 2017


Split mCerulean controlled by inducible araC/pBAD promoter

This a split version of BBa_K2387052. The two halves of mCerulean are fused to leucine zippers to induce the reassembly. Both parts are under the control of the araC/pBad promoter and each part is under the control of a stron RBS. The reassembly process is irreversible. This part can be used as a positive control for experiments using BiFC, as well as a source for the fragments of mCerulean.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Functional Parameters

In order to test the functionality of this split protein under the araC/pBad promoter, the absorbance and the fluorescence spectra of the full length protein and the split protein were measured after induction with 0.002% arabinose (Figure 1). The stability of the shape in both spectra for the full length protein and the split protein show that the split has been successful.

File:T--Wageningen UR--Results Spectra.jpg

Figure 1: Absorbance and fluorescence spectra of full length mCerulean (left) and split mCerulean (right), measured after induction with 0.002% arabinose.

This protein is part of a collection of split proteins developed for BiFC analysis and tested with the CpxR system (BBa_K2387032). These proteins were characterized through multiple experiments using as interacting proteins antiparallel synthetic leucine zippers. The proteins of this collection are the ones found in Table 1.

Table 1: Split Proteins.
Protein Part Number (Full length) Part Number (Split Protein)
mRFP BBa_K2387054 BBa_K2387055
eYFP BBa_K2387003 BBa_K2387065
mVenus BBa_K2387045 BBa_K2387046
sfGFP BBa_K2387047 BBa_K2387048
mCerulean BBa_K2387052 BBa_K2387053