Difference between revisions of "Part:BBa K525222"
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S-layer Corynebacterium halotolerans
 
S-layer Corynebacterium halotolerans
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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_K525222#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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|0.245 h<sup>-1</sup> / 0.109 h<sup>-1</sup>
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|-
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|Doubling time (un-/induced)
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|2.83 h / 6.33 h
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|-
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|}
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</center>
  
 
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Revision as of 21:05, 21 September 2011

S-layer cspB from Corynebacterium halotolerans

S-layer Corynebacterium halotolerans

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) 0.245 h-1 / 0.109 h-1
Doubling time (un-/induced) 2.83 h / 6.33 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 1103
    Illegal XhoI site found at 559
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 226
    Illegal NgoMIV site found at 1315
    Illegal AgeI site found at 217
    Illegal AgeI site found at 458
    Illegal AgeI site found at 505
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 907
    Illegal BsaI.rc site found at 214
    Illegal BsaI.rc site found at 592
    Illegal BsaI.rc site found at 994


Expression in E. coli

The CspB gen was fused with a monomeric RFP (BBa_E1010) using [http://2011.igem.org/Team:Bielefeld-Germany/Protocols#Gibson_assembly Gibson assembly] for characterization.

The CspB|mRFP fusion protein was overexpressed in E. coli KRX after induction of T7 polymerase by supplementation of 0,1 % L-rhamnose 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 CspB and mRFP 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 CspB and mRFP with and without induction. A curve depicting KRX wildtype is shown for comparsion.