Reporter

Part:BBa_K1155007

Designed by: Sylvie Lautru, Anas, Zhou, Damir   Group: iGEM13_Paris_Saclay   (2013-09-10)
Revision as of 02:51, 5 October 2013 by Nad' (Talk | contribs)

RBS-lacZ-term


lacZ gene encodes the protein beta-galactosidase which causes bacteria expressing the gene to appear blue when grown on a medium that contains the substrate analog X-gal.

In order to be easily used by all the other teams, our team improved the biobrick BBa_I732017 designed by iGEM07_USTC by asembly of biobrick BBa_I732017 and B0017.

A composite part constituted of a RBS, full length lacZ gene and a terminator. We used it as one of our reporter gene in Escherichia coli.


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]


Characterization

  • We have tested the Biobricks BBa_K1155007 in the construction: BBa_K1155004 + BBa_K1155007 in pSB1C3

BBa_K1155004 is a promoteur PnirB activated in anaerobic condition by binding of the FNR regulator (see BBa_K1155004 for more information).

What we expected :

+ O2 : FNR inactivated : PnirB inactivated --> lacZ not expressied --> no color

- O2 : FNR activated : PnirB activated --> lacZ expressed --> blue color

What we observed :

We observe blue color in both conditions. This must be because we introduce our construction (BBa_K1155004+BBa_K1155007) in a high copy backbone.

500px

  • We have tested the Biobricks BBa_K1155007 in the construction: BBa_K1155006 + BBa_K1155007 in pSB1K3

BBa_K1155006 is a promoteur PnarK activated in anaerobic condition by binding of the FNR regulator (see BBa_K1155006 for more information).

What we expected :

+ O2 : FNR inactivated : PnirB inactivated --> lacZ not expressied --> no color

- O2 : FNR activated : PnirB activated --> lacZ expressed --> blue color

What we observed :

We observe blue color in anaerobic conditions. We can't do this experience in aerobic conditions.

PsNarKK33108.jpg

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