Fig. 4

(a) pRF_Xp_HPH vector used for ATMT of C. oleaginosus with the five different promoters tested. Elements for plasmid replication and amplification in E. coli include KanR: kanamycin resistance; oriV: plasmid origin of replication; trfA: plasmid replication initiator protein. (b) Growth of C. oleaginosus mutants, with genomically integrated promoter-HPH constructs, and wt cells as controls with and without the addition of hygromycin B. The measurements were performed with four biological replicates. 60Sp, 60 S ribosomal protein L37 promoter; D9FADp, delta-9 fatty acid desaturase promoter; GAPDHp, glyceraldehyde 3-phosphate dehydrogenase promoter; HPH, hygromycin phosphotransferase; Hyg, hygromycin B; PPIp, cyclophilin type peptidyl-prolyl cis-trans isomerase promoter; UBIp, ubiquitin and ubiquitin-like protein promoter; wt, wild type. Plasmid maps were created with Geneious