Part:BBa_K1893019:Experience

Applications of BBa_K1893019

Gp2 was characterised using an arabinose inducible promoter (BBa_K1893016), because constitutive expression, and the resultant arrest of E. coli growth, would result in several potential problems in the cloning and characterisation workflow. The sequence for Gp2 was obtained from the NCBI, and codon optimisation was not required, as the protein is from en E. coli phage.

Characterisation data of Gp2 via BBa_K1893016

Figure 1: Growth inhibition of Top10 cells by induction of Gp2 by L-arabinose. Experiments were performed in E. coli Top10 cell strain cultured at 37°C, which were diluted to 0.05 O.D. and inoculated with L-arabinose at the 0 minute timepoint. O.D. was recorded at 600nm. Reported values represent the mean normalised O.D. for three repeats, with error bars representing standard deviation.

Figure 2: Recovery of growth by TOP10 cells after arabinose-induced pBAD operon was switched off by glucose-mediated catabolite repression. 100uM of L-arabinose was added to the culture 2 hours before the 0 minute timepoint and D-glucose was added at the 0 minute timepoint. pBAD-GFP controls indicate that the pBAD operon was switched off at approximately after approximately 250 minutes. Experiments were performed in E. coli Top10 cell strain cultured at 37°C, which were diluted to 0.05 O.D., which was recorded at 600nm. Reported values represent the mean normalised O.D. for three repeats, with error bars representing standard deviation.

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