Coding
cspB

Part:BBa_K525123

Designed by: Anna Drong   Group: iGEM11_Bielefeld-Germany   (2011-09-11)
Revision as of 18:53, 21 September 2011 by Mlimberg (Talk | contribs) (→‎Expression in E. coli)

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


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 1334
    Illegal XhoI site found at 161
    Illegal XhoI site found at 779
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1313
    Illegal SapI site found at 560
    Illegal SapI site found at 772
    Illegal SapI site found at 1320


Expression in E. coli

For characterization the CspB gen was fused with a monomeric RFP (BBa_E1010) using Gibson assembly.

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 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.
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Categories
//chassis/prokaryote/ecoli
//proteindomain/internal
//rnap/bacteriophage/t7
Parameters
n/aS-layer cspB from Corynebacterium glutamicum with lipid anchor and PT7 and RBS