High Resolution Growth Screen of Ruegeria pomeroyi Transporter Mutants Data September 2021 - June 2022 (C-CoMP Marine Bacterial Transporters project)

High resolution growth screens were used to confirm the phenotype of Ruegeria pomeroyi DSS-3 transporter knockout mutants. Mutants which had demonstrated growth defects on a given substrate as sole carbon source during an initial growth screen were selected and used here. Each mutant was grown on the substrate(s) of interest along side a wildtype analog (pooled-TnSeq library). Growth curves were generated by reading the optical density at 600 nm hourly. The annotations of a transporter's cognate substrate was confirmed when the mutant of said transporter demonstrated significant defect relative to the wildtype analog on the substrate of interest.

本研究通过高分辨率生长筛选实验，对波氏鲁杰氏菌（Ruegeria pomeroyi）DSS-3的转运蛋白敲除突变株的表型进行验证。本次实验选取了在初步生长筛选阶段，于以特定底物为唯一碳源的培养条件下表现出生长缺陷的突变株。每株突变株均与野生型对照（混合转座子测序（pooled-TnSeq）文库）一同在目标底物上培养；通过每小时测定600 nm波长下的光密度值，绘制生长曲线。当某转运蛋白的突变株在目标底物上相较于野生型对照呈现显著生长缺陷时，即可确认该转运蛋白的同源底物注释信息。