Fig. 1: High-throughput conjugative transfer assay of TP114 in broth or on solid media.

a Bacterial conjugation occurring in broth requires Mating Pair Stabilization (MPS) provided by type IVb pili of TP114 to bring cells together and keep them in close contact during plasmid transfer. In this condition, the conjugative pilus and the accessory type IVb pilus will recognize specific molecules on the recipient’s outer membrane. Created in Biorender.com. b Bacterial conjugation on solid media provides high cell density and proximity between donor and recipient cells enabling mating pair formation without the need for MPS via the type IVb pili. A high-throughput conjugation assay of TP114 into ~4000 E. coli single-gene deletion mutants yielded a normal distribution when mating was performed in broth (c) or on solid media (d) when plotting conjugation scores obtained from the ratio of the growth of transconjugants by recipient bacteria (n = 4). e, f Transfer rates of manually validated hits with low, medium, or high conjugation scores. The dotted lines represent the conjugation rates measured with a wild type recipient strain or the average conjugation score obtained in the high-throughput assay. The circles show individual values for each dataset (n = 2).