Difference between revisions of "Part:BBa K525223"
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+ | ===Expression in ''E. coli''=== | ||
+ | The CspB gen was fused with a monomeric RFP ([https://parts.igem.org/Part:BBa_E1010 BBa_E1010]) using [http://2011.igem.org/Team:Bielefeld-Germany/Protocols#Gibson_assembly Gibson assembly] for characterization. | ||
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+ | 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. | ||
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+ | [[Image:Bielefeld_2011_CH3_Growthcurve.png|600px|center|thumb| '''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.''']] | ||
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+ | [[Image:Bielefeld_2011_CH3_RFU_OD.png|600px|center|thumb| '''Figure 2: RFU to OD<sub>600</sub> 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.''']] |
Revision as of 16:28, 21 September 2011
S-layer cspB from Corynebacterium halotolerans with lipid anchor, PT7 and RBS
S-layer cspB from Corynebacterium halotolerans with TAT-Sequence and lipid anchor, PT7 and RBS
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1102
Illegal XhoI site found at 558 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 225
Illegal NgoMIV site found at 1314
Illegal NgoMIV site found at 1425
Illegal AgeI site found at 216
Illegal AgeI site found at 457
Illegal AgeI site found at 504 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 906
Illegal BsaI.rc site found at 213
Illegal BsaI.rc site found at 591
Illegal BsaI.rc site found at 993
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.