Part:BBa_K784011:Experience

Experience of BBa_K784011

This construct has been shown to produce the fluorescent protein mCherry. The plasmid has been transformed into E. coli TOP10 strain. Starters were grown over night, diluted 1:100, grown to ~O.D 0.6 and divided into 48 well plates. IPTG was added (to 1mM concentration) to induce the tac promoter. A control without IPTG was prepared as well. The samples were incubated at 37 degrees for ~3-4.5 hours before measuring fluorescence in the plate reader. The negative control is pSB1C3 with an MCS insert (BBa_K784023). The experiment has been conducted in three plasmid backbones: pSB1C3, pSB3C5 and a custom low copy plasmid backbone notated pCP. Since the Top10 strain has low levels of lacI there was little difference with and without IPTG. The measurements taken in the presence of 1mM IPTG are presented in Table 1.

Table 1: the fluorescence/O.D values measured in the different plasmid backbones

Backbone	Induction time (hours)	Fluorescence/O.D
pSB1C3	4	12831
pSB3C5	4	29818
pCP	3.5	2000

See BBa_K784010 and [http://2012.igem.org/Team:Technion/Project/RS our wiki] for additional info.

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