Figure 1

Reverse engineering cycle for butanol tolerance in S. cerevisiae . The figure represents the evolutionary engineering strategy used in this study. It comprises three phases a) generation of biodiversity by evolutionary engineering and screening for improved isolates (1- and 2-), b) analysis of evolved genomes and identification of genetic variations (SNV: Single Nucleotide Variation, INDELS: INsertion - DELetion and CNV: Copy Number Variation) (3-) and c) reverse genetic engineering of detected variation in a “naive” genetic background and characterization of the engineered strain (5-, 6-). Prioritization of the variations prior reintroduction in naive reference was guided by sequence analysis of offspring from three consecutive back crossing (4-).