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Fig. 2 | Epigenetics & Chromatin

Fig. 2

From: Systematic genetic and proteomic screens during gametogenesis identify H2BK34 methylation as an evolutionary conserved meiotic mark

Fig. 2

Creation of the histone H2A and H2B shuffle strain in the SK1 genetic background. a Creation strategy. Genomic copies encoding H2A (HTA1 and HTA2) and H2B (HTB1 and HBT2) have been deleted and the HTA1 and HTB1 genes are expressed from an autonomous plasmid. The original URA3 plasmid can be replaced by a new HIS3 plasmid with any desired mutation. b Validation of the H2A and H2B shuffle strain. Plasmids encoding Flag-tagged H2A or H2B have been introduced in the strain and these histones have been detected by western blot. They confirm that all genomic copies of H2A/H2B have been deleted and the only H2A/H2B proteins are expressed from the autonomous plasmid in both SK1 and s288c genetic backgrounds. c No growth or sporulation defects are detected in the H2A and H2B shuffle strain in the SK1 background. The genetic manipulation did not affect doubling times during vegetative growth (top) or sporulation efficiency (bottom, see Additional file 1: Figure S1 for more details) when comparing the H2A H2B shuffling strain to a WT or a previously constructed H3 H4 shuffle strain [33]

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