Figure 8

Segregation of the mutations found in the evolved n -butanol tolerant strains IMS0344 and IMS0351 and the F1, F2 and F3 generation of back crossing. The evolved strains IMS0344 and IMS0351 were crossed with IMK439 (MATα isogenic of CEN.PK113-7D and deleted in URA3) to create a diploid. The diploid was sporulated, one haploid segregant with the same tolerance as the evolved strain was selected and the cycle repeated was repeated three times.