Part:BBa_R0011:Experience/iGEM11 Uppsala-Sweden

iGEM11_Uppsala-Sweden: Promoter characterization. We characterised the promotor in a TOP10 E. coli strain together with PcpcG2 (BBa_K592003), PT5lac (BBa_K592008), BBa_J23113, PfixK (BBa_K592006) and as reference promoter we used BBa_J23101. The backbone pSB3K3, the RBS BBa_B0032 and the reporter BFP (BBa_K592100) were used in every construct, and we calculated the relative promoter units, RPU, compared to J23101. The actual test was done in a slightly different way than that presented at the parts.igem. Here we grew the cells in LB medium for 10 hours and we used flow cytometry to quantify the output. The flow cytometer used was a BD FACSAria II, using a 407 nm violet laser and a DAPI filter (band pass 450/40).

The results showed that PLlacO has a strength of 1.9 RPU in TOP10 when repressed by LacI (not lacIq) and a strength of 2.5 RPU when induced with IPTG.