Difference between revisions of "Part:BBa K1088026"

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To repress expression from the lac promoter, the ''lacI'' gene under a constitutive promoter, with a strong RBS and a efficient terminator is placed counter-clockwise to the reporter fusion. The repression can then be relieved with addition of IPTG, which binds and inhibits the function of LacI.  
 
To repress expression from the lac promoter, the ''lacI'' gene under a constitutive promoter, with a strong RBS and a efficient terminator is placed counter-clockwise to the reporter fusion. The repression can then be relieved with addition of IPTG, which binds and inhibits the function of LacI.  
  
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The levels of expression was meassured in ''E. coli'' K-12 MG1655 using fluorescence activated cell sorting (FACS), and proved that their was no expression when grown without IPTG and that there was expression after addition of IPTG. See experience for more details.
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The levels of expression was meassured in ''E. coli'' K-12 MG1655 carrying the part using fluorescence activated cell sorting (FACS). The experiment proved that the reporter fusion wasn't expressed when the strain was grown without IPTG in the media, and that the reporter fusion was expressed after addition of IPTG. See experience for more details.
  
 
This Brick was build to test induction time and IPTG concentration for induction of a similar device (BBa_K1088027) which lacks the linker and GFP.
 
This Brick was build to test induction time and IPTG concentration for induction of a similar device (BBa_K1088027) which lacks the linker and GFP.

Revision as of 21:24, 25 September 2013

B. subtilis dxs-GFP protein fusion (lac promoter with lac inhibitor: IPTG inducible)

This part consist of the dxs gene derived from B. subtilis fused to GFP at the translational level with a 10 AA linker between the proteins. The reporter fusion is under the control of the lac promoter and has a strong RBS.

To repress expression from the lac promoter, the lacI gene under a constitutive promoter, with a strong RBS and a efficient terminator is placed counter-clockwise to the reporter fusion. The repression can then be relieved with addition of IPTG, which binds and inhibits the function of LacI.

The levels of expression was meassured in E. coli K-12 MG1655 carrying the part using fluorescence activated cell sorting (FACS). The experiment proved that the reporter fusion wasn't expressed when the strain was grown without IPTG in the media, and that the reporter fusion was expressed after addition of IPTG. See experience for more details.

This Brick was build to test induction time and IPTG concentration for induction of a similar device (BBa_K1088027) which lacks the linker and GFP.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2460
    Illegal EcoRI site found at 3117
    Illegal PstI site found at 2518
    Illegal PstI site found at 2964
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2460
    Illegal EcoRI site found at 3117
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal PstI site found at 2518
    Illegal PstI site found at 2964
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2460
    Illegal EcoRI site found at 3117
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2460
    Illegal EcoRI site found at 3117
    Illegal PstI site found at 2518
    Illegal PstI site found at 2964
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2460
    Illegal EcoRI site found at 3117
    Illegal PstI site found at 2518
    Illegal PstI site found at 2964
    Illegal NgoMIV site found at 2417
    Illegal AgeI site found at 2310
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2259
    Illegal BsaI.rc site found at 3973
    Illegal SapI.rc site found at 2958