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. Author manuscript; available in PMC: 2018 Jun 8.
Published in final edited form as: Gene. 2012 Nov 29;514(2):82–90. doi: 10.1016/j.gene.2012.11.011

Figure 6. RT-PCR analyses of cDNA products of mRNA transcribed from different supercoiling-reporter plasmids pZXD60, 60A, 61, 62, 63 and 63A in E. coli topA strain VS111 harboring the linear plasmid pZXD51 after 10 min of IPTG induction.

Figure 6

(A) RT-PCR experiments were performed as described under Experimental Procedures. The lower panel is a 1.2% agarose gel containing 1% formaldehyde to show the integrity of the RNA samples used for the RT-PCR experiments. The middle panel is a 12% polyacrylamide gel in 1×TAE buffer to show the PCR products of the cDNA synthesized from 16S rRNA samples isolated from E. coli strain VS111 carrying different supercoiling-reporter plasmids pZXD60, 60A, 62, 63, 63A, 57, and 59 after 10 min of IPTG induction. The upper panel is also a 12% polyacrylamide gel in 1×TAE buffer to show the PCR products of the cDNA samples synthesized from the mRNA samples isolated from E. coli strain VS111 carrying different supercoiling-reporter plasmids pZXD60 (lanes 1 and 4), 60A (lane 5), 62 (lane 2), 63 (lanes 3 and 6), 63A (lane 7), 57 (lane 8), and 59 (lane 9) after 10 min of IPTG induction. (B) Real-time RT-PCR analyses of the mRNA samples for E. coli strain VS111 carrying different supercoiling-reporter plasmids pZXD60, 60A, 61, 62, 63, 63A, and 59 after 10 min of IPTG induction (mean±SD, three independent experiments). Labels: lacZ, the lacZ gene; lacZA, the lacZ gene with an amber mutation in the start codon; lacZR, the lacZ gene in the reverse orientation; GFPuv, the GFPuv gene; GFPuvR, the GFPuv gene in the reverse orientation; tet, the tet gene; tetA, the tet gene with an amber mutation in the start codon.