Part:BBa_K2387053

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