Generator

Part:BBa_K143079

Designed by: Chris Hirst   Group: iGEM08_Imperial_College   (2008-10-27)

Pveg-spoVG GFP expression construct


GFP gene under expresson of the Pveg promoter and spoVG RBS of B. subtilis, with a chloramphenicol resistance marker for ease of selection.


Also contains the 5' and 3' amyE integration sequences to allow integration into B. subtilis at the amyE locus.


This device was used by the Imperial iGEM 2008 team to characterise the Pveg promoter and spoVG RBS (Combined part BBa_K143053) as part of the Biofabricator subtilis project.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2225


Part Characterisation

B.subGFP.PNG

This test construct was used for the characterise the expression output of the promoter pVeg combined with the ribosome binding site spoVG. The characterisation was performed using a plate reader to measure fluorescence and O.D.600. The O.D.600 data was converted to cell number (colony forming units) using the calibration curve O.D.600 vs cell number. Cell number was then used to normalise the fluorescence to give the final normalised units of fluorescence per cell (colony forming unit). The graph below shows the results of a B.subtilis culture containing this test construct and a culture of B.subtilis containing the test construct http://parts.igem.org/Part:BBa_K143082 BBa_K143082] as a negative control. It is clear that B.subtilis containing this test construct is expressing mRFP. To learn more about the characterisation please click this link.

[edit]
Categories
//chassis/prokaryote/bsubtilis
Parameters
None