Difference between revisions of "Part:BBa K525121"
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===Usage and Biology===
 
===Usage and Biology===
 
S-layer proteins can be used as scaffold for nanobiotechnological applications and devices by e.g. fusing the S-layer's self-assembly domain to other functional protein domains. It is possible to coat surfaces and liposomes with S-layers. A big advantage of S-layers: after expressing in ''E. coli'' and purification, the nanobiotechnological system is cell-free. This enhances the biological security of a device.  
 
S-layer proteins can be used as scaffold for nanobiotechnological applications and devices by e.g. fusing the S-layer's self-assembly domain to other functional protein domains. It is possible to coat surfaces and liposomes with S-layers. A big advantage of S-layers: after expressing in ''E. coli'' and purification, the nanobiotechnological system is cell-free. This enhances the biological security of a device.  
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|L-rhamnose for induction of T7 polymerase
 
|L-rhamnose for induction of T7 polymerase
 
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|rowspan="3"|[[Part:BBa_K525305#Purification_of_SgsE_fusion_protein | Purification]]
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|rowspan="3"|[[Part:BBa_K525305#Purification_of_SgsE_fusion_protein | Characteristics]]
 
|Molecular weight
 
|Molecular weight
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K525121 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K525121 SequenceAndFeatures</partinfo>
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===Expression in ''E. coli''===
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The CspB (K525121) 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 auinduction protocol]from promega.
  
  
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K525121 parameters</partinfo>
 
<partinfo>BBa_K525121 parameters</partinfo>
 
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Revision as of 13:31, 21 September 2011

S-layer cspB from Corynebacterium glutamicum with TAT-Sequence and lipid anchor, PT7 and RBS


Usage and Biology

S-layer proteins can be used as scaffold for nanobiotechnological applications and devices by e.g. fusing the S-layer's self-assembly domain to other functional protein domains. It is possible to coat surfaces and liposomes with S-layers. A big advantage of S-layers: after expressing in E. coli and purification, the nanobiotechnological system is cell-free. This enhances the biological security of a device.

This fluorescent S-layer fusion protein is used to characterize purification methods and the S-layer's ability to self-assemble on surfaces.

Important parameters

Experiment Characteristic Result
Expression (E. coli) Localisation cell membrane
Compatibility E. coli KRX
Inductor for expression L-rhamnose for induction of T7 polymerase
Characteristics Molecular weight
Theoretical pI

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1421
    Illegal XhoI site found at 248
    Illegal XhoI site found at 866
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1400
    Illegal SapI site found at 647
    Illegal SapI site found at 859
    Illegal SapI site found at 1407

Expression in E. coli

The CspB (K525121) 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 auinduction protocol]from promega.


Functional Parameters