Investigation of Bar-seq as a method to study population dynamics of Saccharomyces cerevisiae deletion library during bioreactor cultivation

摘要

Schematic depiction of experimental setup for the pooled population dynamics experiments. An aliquot of a pooled haploid prototrophic S. cerevisiae deletion collection [38] was used to inoculate a two stage shake flask seed train in YPD. Growth competition experiments were performed in two sets, each consisting of two seed train stages (Seed 1 and Seed 2) followed by different main cultivation environments. Set 1 (left) varied in vessel architecture that included shake flasks with different culture volumes (SF1 and SF2) and a batch bioreactor (BR). Set 2 (right) varied in cultivation parameter settings that included four fed-batch mode bioreactors (CF4, CF6, DF4 and DF6) with two different feeding modes (CF = constant feed, DF = DO- signal based feed) and two different pH (4 and 6). All cultures were run at 30 °C, the dissolved oxygen (DO) was not controlled. Samples were taken at the end of each seed stage and throughout the bioreactor runs