Composite
SbpA | CFP

Part:BBa_K525406

Designed by: Timo Wolf   Group: iGEM11_Bielefeld-Germany   (2011-09-10)
Revision as of 16:08, 21 September 2011 by JSchwarzhans (Talk | contribs)

Fusion Protein of S-Layer SbpA and mCerulean

Fusion Protein of S-Layer SgsE and mCerulean

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 104
    Illegal BglII site found at 221
    Illegal XhoI site found at 1996
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 76
    Illegal AgeI site found at 3913
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 493
    Illegal BsaI.rc site found at 622


Expression in E. coli

The SbpA (K525401) gen was fused with a mCerulean (BBa_J18930) using [http://2011.igem.org/Team:Bielefeld-Germany/Protocols#Gibson_assembly Gibson assembly] for characterization.

The SbpA|mCerulean fusion protein was overexpressed in E. coli KRX after induction of T7 polymerase by supplementation of 0,1 % L-rhamnose and 1 mM IPTG using the [http://2011.igem.org/Team:Bielefeld-Germany/Protocols/Downstream-processing#Expression_of_S-layer_genes_in_E._coli autinduction protocol] from promega.

Figure 1: Growthcurve of E. coli KRX expressing the fusion protein of SbpA and mCerulean with and without induction. A curve depicting KRX wildtype is shown for comparsion.
Figure 2: RFU to OD600 ratio of E. coli KRX expressing the fusion protein of SbpA and mCerulean with and without induction. A curve depicting KRX wildtype is shown for comparsion.
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Categories
//chassis/prokaryote/ecoli
//function/reporter/fluorescence
//proteindomain/internal
Parameters
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