1. CozE homologs do not dramatically affect cell division in L. plantarum.
CozE homologs do not dramatically affect cell division in L. plantarum. (A) Schematic overview of plasmids harboring sgRNA to knock down expression of cozE homologs lp_1247 (strain IM141) and lp_2217 (strain IM142) and double knockdown of lp_1247 and lp_2217 (strain IM130). The restriction sites utilized for construction of the double-sgRNA plasmids are indicated (see Materials and Methods for details). (B) Phase-contrast micrographs of cells depleted of lp_1247 or lp_2217 or both. Cells were grown in the presence of 25 ng/ml SppIP. The scale bar is 5 µm. (C) Cell length analysis of the corresponding strains compared to control cells (strain IM133). Cell lengths were measured using MicrobeJ (59). Cells were split in four groups based on lengths, and the proportion of cells in each group is plotted. Numbers of cells analyzed were as follows: n = 204 for control cells (strain IM133), n = 256 for sgRNA-lp_1247 (strain IM141), n = 190 for sgRNA-lp_2217 (strain IM142), and n = 244 for sgRNA-lp_1247-lp_2217 (strain IM130). Plus signs (+) indicate that cells were grown in the presence of 25 ng/ml SppIP. (D) The transcription levels of genes lp_1247 and lp_2217 in different strains as analyzed by droplet digital PCR. In addition to lp_2217 cells (strain IM142), lp_1247 cells (strain IM141), and the double-knockdown cells (strain IM130), control cells with nontargeting sgRNA (strain IM133) were included in the analysis.
Ine Storaker Myrbråten et al. mSphere 2019; doi: /mSphere