Part:BBa_K525223

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:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1102
  Illegal XhoI site found at 558
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE 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
  
• 1000
  INCOMPATIBLE 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.

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.