Measurement

Part:BBa_K862000

Designed by: Charlotte Bunne, Mariam Harmouche, Stefan Holderbach, Anna Huhn and Jakob Kreft (in alphabetical order)   Group: iGEM12_Heidelberg_LSL   (2012-06-08)
Revision as of 12:26, 25 June 2012 by Dniopek (Talk | contribs)

precA-LacZ-doubleTerminator

This is a part for the precise quantification of UV-radiation or radioactive radiation in E. coli (recA+) strains, i.e. BL21(DE3). It consits of a recA Promoter (part BBa_J22106) fused to a LacZ reporter cloned in front of a double terminator (part BBa_K173004).


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
    COMPATIBLE WITH RFC[1000]



Characterisation

This part was characterized during the 2012 HS competition by the Heidelberg_LSL team

Fig. 1: Comparison of parts BBa_K862000 (precA_LacZ) and BBa_K862001 (psulA_LacZ) via ONPG assay of Bl21(DE3) UV-irradiated for different times. Both constructs show a strong correlation between the UV-irradiation time and the LacZ activity (as measured by the production of o-nitrophenol).



Fig. 2: X-gal assay of Bl21(DE3) transformed with BBa_K862000 (precA_LacZ), BBa_K862001 (psulA_LacZ) and BBa_K862002 (precB_LacZ) and UV-irradiated for different times. All constructs show a strong positive correlation between UV induction time and coloring of the wells. PrecA-LacZ (BBa_K862000) gives the lowest reporter background expression whereas psulA-LacZ (BBa_K862001) gives the highest overall coloring of the samples.

[edit]
Categories
//chassis/prokaryote
//chassis/prokaryote/ecoli
//function/sensor
Parameters
None