Difference between revisions of "Part:BBa K2235003"
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<partinfo>BBa_K2235003 short</partinfo>
 
<partinfo>BBa_K2235003 short</partinfo>
  
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This composite part encodes all the necessary regulatory elements for a functional chemically inducible system, the cumate system. In particular, there is an expression cassette encoding the reporter gene, mTagBFP, <partinfo>K592100</partinfo> and there are two additional multiple cloning sites (MCS) flanking the reporter (check the design page for more information). The repressor of the system, CymR, is located upstream of the expression cassette and it is constitutively expressed. The RNA spinach is an additional reporter protein following CymR aiming to detect its expression upon addition of its cognate fluorophore.
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This composite part encodes all the necessary regulatory elements for a functional chemically inducible system, the cumate system. In particular, there is an expression cassette encoding the reporter gene, mTagBFP, <partinfo>K592100</partinfo> and there are two additional multiple cloning sites (MCS) flanking the reporter (check the design page for more information). The repressor of the system, CymR <partinfo>K415202</partinfo> , is located upstream of the expression cassette and it is constitutively expressed. The RNA spinach, <partinfo>K1330000</partinfo>, is an additional reporter protein following CymR aiming to detect its expression upon addition of its cognate fluorophore.
  
  

Revision as of 12:53, 23 October 2017


Cumate gene switch expressing BFP

This composite part encodes all the necessary regulatory elements for a functional chemically inducible system, the cumate system. In particular, there is an expression cassette encoding the reporter gene, mTagBFP, BBa_K592100 and there are two additional multiple cloning sites (MCS) flanking the reporter (check the design page for more information). The repressor of the system, CymR BBa_K415202 , is located upstream of the expression cassette and it is constitutively expressed. The RNA spinach, BBa_K1330000, is an additional reporter protein following CymR aiming to detect its expression upon addition of its cognate fluorophore.


Usage and Biology

The cumate system have been previously described as a versatile and tightly regulated expression system induced by 4-isopropylbenzoic acid (cumate) in various strains of E.coli (1). It can be employed for the regulation and production of heterologous proteins in a dose-dependent manner. Moreover, the induction is achieved quickly within minutes therefore, a robust and homogenous expression of any protein can be achieved upon addition of cumate.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 418
    Illegal XhoI site found at 199
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 101

References

1. Choi Y, Morel L, Le Francois T, Bourque D, Bourget L, Groleau D et al. Novel, Versatile, and Tightly Regulated Expression System for Escherichia coli Strains. Applied and Environmental Microbiology. 2010;76(15):5058-5066.