Part:BBa_K1155007
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.
- 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 could not do this experience in aerobic conditions.
TO CONCLUDE : Our biobrick BBa_K1155007 works very well because we can observe blue colonies on Xgal. It can be easily used as promoter expression rapporter
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