posted on 2022-01-05, 18:35authored byYing Li, Adrien Boes, Yuanyuan Cui, Shan Zhao, Qingzhen Liao, Han Gong, Eefjan Breukink, Joe Lutkenhaus, Mohammed Terrak, Shishen Du
Plasmids pDSW208, pSEB429 (pDSW08, P204::ftsW) or its derivatives harboring a dominant-negative ftsW mutation were transformed into strain SD237 [W3110, leu::Tn10 ftsW::kan/pDSW406 (pBAD33, PBAD::ftsW)] on LB plates with ampicillin and 0.2% arabinose. The next day, a single transformant of each resulting strain was resuspended in 1 ml of LB medium, and serially diluted ten-fold. 3 μl of each dilution was spot on LB plates with antibiotics, with or without increasing concentrations of IPTG. Plates were incubated at 30°C overnight and imaged.