Figure 5.

Plasmid shuffle used to generate strains expressing human cDNAs with missense mutations. Yeast haploid knockout strains covered by wild-type human cDNAs on URA3-marked vectors were transformed with the following LEU2-marked vectors (empty, wild-type human cDNA, and human cDNA with missense mutation) and maintained on −Ura −Leu media. Strains were plated on −Leu +5-FOA media to generate haploid yeast knockouts covered by LEU2-marked vectors. Strains were confirmed to have lost the URA3-marked plasmid by streaking on −Ura media to observe no growth. In the presented example, hSsrp1-K33E is able to complement pob3∆, but S481P results in a nonfunctional hSsrp1 protein.