Fig. 7: Integration of a 151 kb E. coli chromosomal region into the V. natriegens chromosome.

a Scheme of the swap and conjugational transfer of the 151 kb MFDdel4 chromosomal region from E. coli to V. natriegens and subsequent integration. Cell colours represent the corresponding editing step. b Shown are agarose gels of PCRs from the transfer and integration of a 151 kb chromosomal region from E. coli into the recJ locus of V. natriegens. PCRs were done for the acceptor strain (V. natriegens) and for the edited strain (V. natriegensrecJ::MFDdel4). PCRs “1–6” result in the amplification of evenly distributed fragments of the E. coli del4 region within the V. natriegens chromosome. PCR “7” results in the amplification of a 4479 bp fragment, if the recJ locus is unchanged. PCR “8” and “9” amplify the transition of the del4 region to the adjacent chromosomal regions. PCR was repeated once with identical results. Chromosomal overviews including homologous regions (Hα, Hβ of each PCR are shown above each agarose gel. c Shown is the next-generation sequencing (NGS) coverage of the edited strain V. natriegensrecJ::MFDdel4 (light grey) compared to the reference V. natriegens strain (dark grey). Both NGS reads were aligned against the edited reference genome (recJ::MFDdel4). The sector of the del4 insertion as well as the complete genome coverage (circle) are shown. Reads of the complete del4 insertion (between dashed lines) are present for the edited strain, while no reads are present for the wild-type strain (red rectangle).