Difference between revisions of "Part:BBa K1155007"

Latest revision as of 07:58, 5 October 2013

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

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