Difference between revisions of "Part:BBa K525406"
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<partinfo>BBa_K525406 short</partinfo>
 
<partinfo>BBa_K525406 short</partinfo>
  
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Fusion Protein of S-Layer SgsE and mCerulean  
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Fusion Protein of S-Layer SgsE and mCerulean
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===Important parameters===
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<center>
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{|{{Table}}
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!Experiment
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!Characteristic
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!Result
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|-
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|rowspan="5"|[[Part:BBa_K525405#Expression_in_E._coli | Expression (''E. coli'')]]
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|Localisation
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|Inclusion body
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|-
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|Compatibility
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|''E. coli'' KRX and BL21(DE3)
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|-
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|Induction of expression
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|Expression of T7 polymerase + IPTG, lactose
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|-
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|Specific growth rate (un-/induced)
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|0.116 h<sup>-1</sup> / 0.092 h<sup>-1</sup>
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|-
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|Doubling time (un-/induced)
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|5.95 h / 7.51 h
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|-
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|}
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</center>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 20:11, 21 September 2011

Fusion Protein of S-Layer SbpA and mCerulean

Fusion Protein of S-Layer SgsE and mCerulean

Important parameters

Experiment Characteristic Result
Expression (E. coli) Localisation Inclusion body
Compatibility E. coli KRX and BL21(DE3)
Induction of expression Expression of T7 polymerase + IPTG, lactose
Specific growth rate (un-/induced) 0.116 h-1 / 0.092 h-1
Doubling time (un-/induced) 5.95 h / 7.51 h

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.