Difference between revisions of "Part:BBa K1073035:Experience"

iGEM Team Braunschweig 2013

Using ampicillin containing complex medium, growth of cells containing this device was induced by adding synthetic autoinducer N-buturyl homoserine lactone to the culture broth. The required transcription regulator RhlR is encoded in the device. Cells also exhibit a bright pink color due to the constitutively expressed eforRed chromoprotein. Thus eforRed was used as a selection marker identifying cells containing this device. 2xYT medium containing ampicillin was used for cultivation and cells were incubated at 37°C and 250 rpm.

iGEM Team Braunschweig 2013: Comparison of growth curves of E. coli Top10 containing BBa_K1073035 on the high copy plasmid pSB1C3 in the presence and absence of autoinducer N-buturyl-HSL in the culture broth.

Due to the leakiness of the las promoter the beta lactamase gene ampR is also expressed to some extend when no N-buturyl-HSL is added to the culture broth. However, expression is much higher when the autoinducer is added.

Growth of cells could also be induced when culture supernatant of BBa_K1073034 containing N-buturyl-HSL synthesized by RhlI encoded in BBa_K1073034 is added to cultures on agar plates containing ampicillin.

iGEM Team Braunschweig 2013: Growth of E. coli Top10 containing BBa_K1073035 on the high copy plasmid pSB1C3 induced by the supernatant of expressed BBa_K1073034. The supernatant contains N-buturyl-HSL produced by RhlI. A piece of filter paper was soaked with supernatant of a BBa_K1073034 and placed on the agar plate containing ampicillin (0.29 mM).