Fig. 2

Functional analysis of the gene set1 of M. lusitanicus. a Scheme of mutant locus after set1 locus replacement on the genome of wild-type strain by the pyrG construct by homologous recombination. The recombination sites on the genome are marked with dashed lines. The binding sites of oligonucleotides used to check genic deletion and the size of amplified fragments are denoted on the diagram. b PCR products of set1 locus of wild-type strain MU636 (WT) and selected set1 mutants were amplified with the primers Set1-UF and Set1-DR (Additional file 1: Table S2) and separated by electrophoresis. The expected band (black arrows) for the mutant and wild-type locus corresponded to 5.1 kb and 3.1 kb, respectively. c H3K4 methylation in the wild-type strain (WT) and set1 mutants was determined by western blot analysis by using specific antibodies against H3K4 mono- (me1), di- (me2) and trimethylation (me3). As a control, S. cerevisiae protein extract was included. The protein samples were also incubated with anti-histone H3 antibody as a loading control