Results
Contents |
NB: unless differently specified, all tests were performed in E. coli MGZ1 in M9 supplemented medium at 37°C. For the cloning of the parts, E. coli TOP10 was used.
Parts assembly
Characterization of basic modules
Characterization of promoters pTet and pLux
Characterization of enzymes AiiA and LuxI
Characterization of the efficiency of RBSs from the community collection
Identification of bacterial growth parameters
The bacterial growth curve has been modelled as a logistic curve and is represented by the following equation:
dN/dt=N*μ*(Nmax-N)/Nmax
N(0)=n0
where μ represents the growth rate of the cells (E. coli MGZ1 in M9 supplemented medium) and Nmax represents the maximum number of cells in the well. For a detailed description of the parameters, see modelling section. For details on parameters identification, see identification section.
The growth curves in all the performed experiments are measured in O.D.600. Since the N species in the model is expressed in cell number, a conversion factor has been estimated. The conversion factor KO.D.toC.F.U. has been estimated as follows.
in fresh M9 medium. These cultures were grown for further 1 hour at 37°C, 220 rpm.
NB: from now on, cultures must be placed in ice to stop cell growth. Culture O.D.600 TECAN O.D.600 Spectrophotometer C.F.U. Dilution Factor (10^) N C1 0,367950002 0,758004416 990 5 990000000 C1 0,044649998 0,091982322 141 6 1410000000 C1 0,004799999 0,009888356 136 5 136000000 C1 0,000549998 0,001133037 20 6 200000000 C2 0,54840003 1,129744917 165 4 16500000 C2 0,058200002 0,119896339 23 5 23000000 C2 0,008700002 0,017922652 251 3 2510000 C2 0,000100002 0,000206011 24 4 2400000