Fig. 9: Transfer of the RP4 conjugation system to D. dadantii and transfer of the β-galactosidase GanB ORF to E. coli DH5α.
From: A multifunctional system for genome editing and large-scale interspecies gene transfer
a Scheme of the swap and conjugational transfer of the RP4 conjugation system from E. coli MFDpir to D. dadantii and subsequent integration. Cell colours represent the corresponding editing step. b Shown is the next-generation sequencing (NGS) coverage (reads/base) of D. dadantii lacZ::RP4 (light grey) and D. dadantii (dark grey) at the RP4 insert locus mapped against the D. dadantii lacZ::RP4 reference genome. c Conjugation test of pSwap from D. dadantii lacZ::RP4 to E. coli DH5α. Controls for donor and acceptor strains are indicated. d Scheme of the swap and conjugational transfer of the ganB ORF from D. dadantii (D.d.)lacZ::RP4 to E. coli DH5α and subsequent integration. Cell colours represent the corresponding editing step. e PCR verification of the Drop of D. dadantii ganB ORF in the E. coli DH5α lacZω ORF. Two clones after Drop compared to DH5α. PCR was repeated once with identical results. f Blue colony phenotype of DH5α lacZω::ganB upon induction with IPTG and X-gal. The pDrop plasmid is still present in some colonies of the plated non-selective overnight culture resulting in red colonies.
