Difference between revisions of "Part:BBa K525406"
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|Localisation | |Localisation | ||
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===Expression in ''E. coli''=== | ===Expression in ''E. coli''=== | ||
The SbpA (K525401) gen was fused with a mCerulean ([https://parts.igem.org/Part:BBa_J18930 BBa_J18930]) using [http://2011.igem.org/Team:Bielefeld-Germany/Protocols#Gibson_assembly Gibson assembly] for characterization. | The SbpA (K525401) gen was fused with a mCerulean ([https://parts.igem.org/Part:BBa_J18930 BBa_J18930]) using [http://2011.igem.org/Team:Bielefeld-Germany/Protocols#Gibson_assembly Gibson assembly] for characterization. | ||
Revision as of 20:23, 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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 104
Illegal BglII site found at 221
Illegal XhoI site found at 1996 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 76
Illegal AgeI site found at 3913 - 1000INCOMPATIBLE 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.
