Composite
SgsE | RFP

Part:BBa_K525304

Designed by: Timo Wolf   Group: iGEM11_Bielefeld-Germany   (2011-09-10)
Revision as of 15:35, 21 September 2011 by JSchwarzhans (Talk | contribs) (→‎Expression in E. coli)

Fusion Protein of S-Layer SgsE and mCherry RFP

Fusion Protein of S-Layer SgsE and mCherry RFP

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 167
    Illegal BglII site found at 1022
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 76
    Illegal AgeI site found at 3112
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1657


Expression in E. coli

The SgsE (K525304) gen was fused with a mCherry RFP (BBa_J18932) using [http://2011.igem.org/Team:Bielefeld-Germany/Protocols#Gibson_assembly Gibson assembly] for characterization.

The SgsE|mCherry RFP 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 Sgse and mCherry RFP 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 Sgse and mCherry RFP 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
None