Difference between revisions of "Part:BBa K525234"

 
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__NOTOC__
 
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<partinfo>BBa_K525234 short</partinfo>
 
<partinfo>BBa_K525234 short</partinfo>
  
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Fusion Protein of mRFP and S-layer cspB from Corynebacterium halotolerans with TAT-sequence, PT7 and RBS
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Fusion protein of mRFP and S-layer CspB (PS2) from ''Corynebacterium halotolerans'' with TAT-sequence, T7 promoter and RBS.
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S-layers (crystalline bacterial surface layer) are crystal-like layers consisting of multiple protein monomers and can be found in various (archae-)bacteria. They constitute the outermost part of the cell wall. Especially their ability for self-assembly into distinct geometries is of scientific interest. At phase boundaries, in solutions and on a variety of surfaces they form different lattice structures. The geometry and arrangement is determined by the C-terminal self assembly-domain, which is specific for each S-layer protein. The most common lattice geometries are oblique, square and hexagonal. By modifying the characteristics of the S-layer through combination with functional groups and protein domains as well as their defined position and orientation to eachother (determined by the S-layer geometry) it is possible to realize various practical applications ([http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.2006.00573.x/full Sleytr ''et al.'', 2007]).
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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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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This fluorescent S-layer fusion protein is used to characterize purification methods and the S-layer's ability to self-assemble on surfaces.
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===Important parameters===
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<center>
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{|{{Table}}
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!Experiment
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!Characteristic
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!Result
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|-
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|rowspan="3"|[[Part:BBa_K525232#Expression_in_E._coli | Expression (''E. coli'')]]
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|Localisation
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|Inclusion body
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|-
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|Compatibility
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|''E. coli'' KRX and BL21(DE3)
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|-
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|Inductor for expression
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|T7 polymerase
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|-
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|rowspan="3"|[[Part:BBa_K525405#Purification_of_SbpA_fusion_protein | Purification]]
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|Molecular weight
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|137.0 kDa
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|-
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|Theoretical pI
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|4.93
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|-
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|Excitation / emission
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|584 / 607 nm
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|-
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|}
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</center>
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<partinfo>BBa_K525234 parameters</partinfo>
 
<partinfo>BBa_K525234 parameters</partinfo>
 
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===Expression in ''E. coli''===
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===Intracellular localisation===
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===Purification===

Revision as of 14:37, 20 September 2011

Fusion Protein of mRFP, S-layer cspB from Corynebacterium halotolerans with TAT-sequence, PT7, RBS

Fusion protein of mRFP and S-layer CspB (PS2) from Corynebacterium halotolerans with TAT-sequence, T7 promoter and RBS.

S-layers (crystalline bacterial surface layer) are crystal-like layers consisting of multiple protein monomers and can be found in various (archae-)bacteria. They constitute the outermost part of the cell wall. Especially their ability for self-assembly into distinct geometries is of scientific interest. At phase boundaries, in solutions and on a variety of surfaces they form different lattice structures. The geometry and arrangement is determined by the C-terminal self assembly-domain, which is specific for each S-layer protein. The most common lattice geometries are oblique, square and hexagonal. By modifying the characteristics of the S-layer through combination with functional groups and protein domains as well as their defined position and orientation to eachother (determined by the S-layer geometry) it is possible to realize various practical applications ([http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.2006.00573.x/full Sleytr et al., 2007]).


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 Inclusion body
Compatibility E. coli KRX and BL21(DE3)
Inductor for expression T7 polymerase
Purification Molecular weight 137.0 kDa
Theoretical pI 4.93
Excitation / emission 584 / 607 nm


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 799
    Illegal PstI site found at 1125
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 799
    Illegal PstI site found at 1125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1876
    Illegal XhoI site found at 1332
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 799
    Illegal PstI site found at 1125
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 799
    Illegal PstI site found at 1125
    Illegal NgoMIV site found at 999
    Illegal NgoMIV site found at 2088
    Illegal AgeI site found at 87
    Illegal AgeI site found at 680
    Illegal AgeI site found at 792
    Illegal AgeI site found at 990
    Illegal AgeI site found at 1231
    Illegal AgeI site found at 1278
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1680
    Illegal BsaI.rc site found at 987
    Illegal BsaI.rc site found at 1365
    Illegal BsaI.rc site found at 1767


Expression in E. coli

Intracellular localisation

Purification