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Fig. 3 | BMC Microbiology

Fig. 3

From: Functional characterization of thiolase-encoding genes from Xanthophyllomyces dendrorhous and their effects on carotenoid synthesis

Fig. 3

PCR analyses of S. cerevisiae haploid strains. S. cerevisiae strains Sc-ERG10sc (carrying plasmid YEpNP-10sc) and Sc-cERG10xd (carrying plasmid YEpNP-c10xd) were analyzed by PCR to confirm the expected genotype. As controls, X. dendrorhous UCD 67–385 strain (Lane 1), S. cerevisiae strain S288c (Lane 2), S. cerevisiae diploid strain Sc-ERG10/erg10 (Lane 3) and a no-template control (Lane 6), were included. S. cerevisiae Sc-ERG10sc haploid strain (Lane 4) and S. cerevisiae Sc-cERG10xd haploid strain (Lane 5) were analyzed to assess: a absence of chromosomal ERG10 from S. cerevisiae (primers erg10scF and erg10scDWR); b presence of geneticin resistance cassette in S. cerevisiae (primers KanMXF2 and KanMXR2); c presence of ERG10 from S. cerevisiae (primers erg10scF and erg10scR); and d presence of ERG10 from X. dendrorhous (primers Thio2Fw and Thio2Rv). The molecular size markers Lambda DNA/HindIII (Lane M; 23.1, 9.4, 6.6, 4.4, 2.3, 2 and 0.6 kb) and GeneRuler 1 kb Plus (Lane 1kB, band size in kb is indicated) were used. On the right side of the picture, a schematic diagram of the amplification products is included; arrows represent primer sets with a letter indicating in which panel they were used. UP and DOWN (in blue) correspond to chromosomal regions located approximately 300 bp upstream and downstream of the S. cerevisiae ERG10 gene, respectively, KanMX4 corresponds to the geneticin (G418) resistance module, and pACT4 (in red) and tTDH3 (in green) correspond to promoter and terminator regions in the vector YEpNP, respectively

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