Fig. 2.
Standard stability tests of constructed vectors. E. coli DH5α transformants were grown overnight with antibiotic (point 0) and for 40 generations without antibiotic. Every 20 generations appropriate dilution was plated on L agar to obtain approximately 100 colonies. The colonies were replica plated to test for chloramphenicol resistance. Plasmid retention was expressed as percentage of CmR colonies. The results shown are average from three experiments with standard deviation. a Retention of constructed vectors. b Plasmid copy number estimated by RealTime qPCR. Plasmid copy relative to the chromosome was assayed in three independent biological samples with three technical replicates each. Average results for plasmids are presented with SD as follows: 0.06; 1.75; 0.21; 0.16; 0.95; 0.32; 0.17; 6.96, respectively. c Stabilizing properties of active partitioning operon from RA3 in pABB32 and pABB35 vectors. d Effect of IPTG-induced expression of P. aeruginosa parA-parB operon on pABB34 plasmid retention. DH5α(pABB34) cultures were grown in L broth with various concentrations of IPTG