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. 2022 Sep 23;8(38):eabq0304. doi: 10.1126/sciadv.abq0304

Fig. 6. Involvement of A. loripes Cys1 and Cys2 proteins in cysteine biosynthesis in vivo.

Fig. 6.

Yeast spot assays in S. pombe mutants grown in the presence or absence of l-cysteine for l-cysteine auxotrophy gene complementation with l-leucine as selection marker. (A) A. loripes Cys1a gene (Acys1a) complementation assay. (B) A. loripes Cys2 gene (Acys2) complementation assay. Strains: WT (ED666), ade6 ura4 leu1; cys1aΔ, ade6 ura4 leu1 cys1a::kanMX4; and cys2Δ, ade6 ura4 leu1 cys2::kanMX4. All assays were carried out with the addition of adenine and uracil as the background strain (WT) is auxotrophic for both. The plasmid pREP3xN, containing LEU2 as selection marker to complement leucine auxotrophy in the ED666 WT strain, was used as the expression vector and transformed empty, with A. loripes Acys1a gene or with A. loripes Acys2 gene. All assays were grown in EMM medium and done in triplicates, with the figure showing one representative assay.