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Fig. 2 | Microbial Cell Factories

Fig. 2

From: SCRaMbLE generates evolved yeasts with increased alkali tolerance

Fig. 2

Characterization of SCRaMbLEd strains with increased alkali tolerance. a Characterization of lethality for SCRaMbLE system. Four strains (synV strain carrying pYW079, synV carrying pYW180, synV&X carrying pYW079 and synV&X carrying pYW079) were cultured in SC-Ura medium and exposed in β-estradiol for varied time (2 h, 4 h, 6 h, 8 h, 12 h, 16 h), and then diluted on YPD plates. SynV and synV&X strains containing empty vector pRS416 were used as control strains. b SCRaMbLEd synV&X strains on YPD medium and selective medium (YPD medium at pH 8.0). Strains on YPD medium were cultured 2 days and strains on selective medium were cultured 4 days in 30 °C before photographed. SynV&X strain containing empty vector pRS416 was used as a control strain. c Phenotype verification of SCRaMbLEd strains. SynV and synV&X strains were used as control strains. d PCRTag analysis. PCRTags of genes YER042W, YER161C, YER163C and YER182W were labeled in red, indicating deletions of these regions in strain yML008. None amplification of PCRTags of YEL017W, YER010C, YER081W, YER118C in both synV and yML008 strains was caused by nonspecific primers. SynV strain was used as a control strain. All PCRTag primers were listed in Additional file 1: Table S1

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