Additional file 2 of Discovery of a novel translation-machinery-associated protein that positively correlates with cellulase production

Supplementary material 2: Figure S2 Interaction analysis between PoTma15 and PoXlnR using yeast two-hybrid assay. The open reading frame (ORF) of PoTma15 was cloned into the plasmid pGBKT7 containing the GAL4 DNA binding domain (DNA-BD). The ORF of the transcription factors PoXlnR was amplified using cDNA from P. oxalicum WT as the template and cloned into the plasmid pGADT7 containing the GAL4 activation domain (AD). The plasmid with AD (AD-PoxlnR) was transformed into yeast Y187 to obtain the Y187-AD-XlnR strain. The plasmid with BD (BD-PoTma15) was transformed into yeast Y2H Gold-BD to obtain the Y2H Gold-BD-Tma15 strain. (A) The test of toxicity and autoactivation of the Y187-AD-XlnR strain (left) and Gold-BD-Tma15 strain (right). The results demonstrated that there is no toxicity and autoactivation. (B) The hybridized strains were tested on Quadruple Dropout (QDO, QDO–Ade/–His/−Leu/−Trp) with X-α-gal, where the positive control colony turned blue, and the negative control colony was kept white. The results demonstrated no direct interaction between PoTma15 and PoXlnR.