Figure 1.
Deletion construction and gene dispensability. (a) Gene deletion cassette containing the KanMX4 gene flanked by unique barcodes (UPTAG/DNTAG) and regions of homology to the gene of interest (RHG). The cassette replaced the ORF of interest by homologous recombination at the RHG regions. (b) Construction of deletion mutants. All 4,836 protein coding genes were deleted using serial extension PCR (31.3%), block PCR (63.2%), or total gene synthesis (5.4%). The remaining 78 genes could not be confirmed as deleted due to ambiguous sequencing results, recombination failure, or inviability of the heterozygous diploids. (c) Dispensability of 4,836 protein coding genes. For 3,626 (2,729+897) genes the dispensability was previously unknown. ND=not done.