Skip to main content
Fig. 6 | Microbial Cell Factories

Fig. 6

From: Whole transcriptome analysis and gene deletion to understand the chloramphenicol resistance mechanism and develop a screening method for homologous recombination in Myxococcus xanthus

Fig. 6

Integration of homologous recombination by Cm selection in M. xanthus DK-2566. a Schematic diagram of the integration into the genome of DK-2566 via regular homologous recombination by pR6K-Cm-11. Cm-positive clones were further investigated by PCR with primers (marked in P1 and P2) designed onto the Cm resistant gene CmR (shown in orange). b Schematic diagram of the irregular integration into the genome of DK-2566 via irregular homologous recombination by pR6K-Cm-11. Firstly, suicide plasmid pR6K-cm-11 was electro-transformed into the cell of DK-2566. c Identification of integration into the genome of DK-2566 by agarose gel electrophoresis of PCR products in Cm-positive clones. Lane M1-1,-5,-6,-7, M2-5, -6, -7, -8, -9, -10, -11, M3-8 show 0.9 kb bands showing CmR gene of strains DK-2566-11 by primer P1 and P2. No band in other lanes was amplified, although these clones can grow in a Cm agar plate. d Identification of correct integration into the genome of DK-2566 was performed using agarose gel electrophoresis of PCR products of Cm-positive clones. Primers (marked in P3 and P4) were designed to target both ends of homologous arm (shown in light green). Lanes 1–7 show 1.8 kb bands of the vector region integrated into the genome in strain DK-2566-11 by primer pairs P3 and P4. e Verification of irregular integration occurring at the region of the vector was assessed by agarose gel electrophoresis. Primers pairs P5 and P6 located in the backbone were designed to verify that the recombination did not occur at the vector region. f Chromatograms of the 1.8 kb PCR product amplified and sequenced using primers P3 and P4 as sequencing primers

Back to article page