Part:BBa_K1155004:Experience

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Applications of BBa_K1155004

We have tested the activity of BBa_K1155004 by observing the expression of lacZ.

Construction: BBa_K1155004 + BBa_K1155007 in pSB1C3 ( construction obtained not yet sequenced)

Strains were plated on LB + Xgal and incubated in aerobic or anaerobic conditions.

What we expected :

+ O2 : FNR inactivated : PnirB repressed --> no color

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

What we observed :

+ O2 : --> blue color (plate on the left)

- O2 : --> blue color (plate on the right)

These results indicate that the PnirB (BBa_K1155004) is activated in both aerobic and anaerobic conditions. So the PnirB is actually well induced by FNR but there is some issues with the repression. Our hypothesis is that as the pSB1C3 is a high copy number plasmid, the residual activity PnirB is sufficiant to observe a blue color even if this promoter is not activated by FNR binding. The stability of beta-galactosidase can also be an explanation. We should test these construction in low copy number plasmid like pSB1K3 or with another rapporteur gene.

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