Journal: Microbial Cell Factories

Article Title: Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae

doi: 10.1186/s12934-015-0223-7

Figure Lengend Snippet: Testing promoters for differential expression on sucrose. SUC2, GLC3, MAL12 , and GPH1 promoters were used to control GFP expression. Each strain was grown to mid-exponential phase on minimal medium containing 10 g/L glucose or 10 g/L sucrose prior to flow cytometry-based GFP measurement. Mean GFP fluorescence levels in arbitrary units (au) from duplicate fermentations with error bars representing the standard deviation are shown.

Article Snippet: Plasmids P TEF1 -yEGFPCLN2PEST-pRS406, P SUC2 -yEGFPCLN2PEST-pRS406, and P SUC2 -GFPantisense-CYC1t-pRS413 will be made available from AddGene ( www.addgene.org/ ).

Techniques: Quantitative Proteomics, Control, Expressing, Flow Cytometry, Fluorescence, Standard Deviation

Journal: Microbial Cell Factories

Article Title: Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae

doi: 10.1186/s12934-015-0223-7

Figure Lengend Snippet: Tuning the timing of gene expression using different ratios of glucose to sucrose. A strain expressing the destabilized GFP gene driven by the SUC2 promoter was grown in media containing the indicated concentrations of glucose and sucrose. Extracellular glucose, sucrose, and fructose concentrations were measured using HPLC during the initial growth phase alongside GFP expression levels. Population density (OD 660nm ) and GFP expression levels were measured up to 56 hours of shake-flask cultivation. All data points and error bars represent the mean and standard deviation from triplicate cultivations.

Article Snippet: Plasmids P TEF1 -yEGFPCLN2PEST-pRS406, P SUC2 -yEGFPCLN2PEST-pRS406, and P SUC2 -GFPantisense-CYC1t-pRS413 will be made available from AddGene ( www.addgene.org/ ).

Techniques: Gene Expression, Expressing, Standard Deviation

Journal: Microbial Cell Factories

Article Title: Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae

doi: 10.1186/s12934-015-0223-7

Figure Lengend Snippet: Comparison of TEF1 and SUC2 promoter strengths. GFP fluorescence (a) and population density (b) were measured for P TEF1 - GFP and P SUC2 - GFP expressing strains in 1% glucose, 1% sucrose containing medium over 48 hours. Mean and standard deviation for triplicate cultivations are shown.

Article Snippet: Plasmids P TEF1 -yEGFPCLN2PEST-pRS406, P SUC2 -yEGFPCLN2PEST-pRS406, and P SUC2 -GFPantisense-CYC1t-pRS413 will be made available from AddGene ( www.addgene.org/ ).

Techniques: Comparison, Fluorescence, Expressing, Standard Deviation

Journal: Microbial Cell Factories

Article Title: Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae

doi: 10.1186/s12934-015-0223-7

Figure Lengend Snippet: Dynamic repression of GFP expression using sucrose mediated RNAi. (a) The expression of an antisense RNA results in the destruction of complementary mRNA via Dicer and Argonaute enzymes. (b) Expression of the full GFP ORF in the antisense direction is triggered during growth on sucrose using the SUC2 promoter, causing constitutively regulated ( TEF1 promoter) GFP expression to be repressed via Dicer/Argonaute-mediated RNA interference. GFP expression levels (c) , and population density (d) were measured for P TEF1 - GFP expressing strains both with (‘RNAi’, black triangles) and without (‘control’, green circles) a P SUC2 - GFP antisense construct. The GFP expression level from the control strain was set to ‘100%’, with GFP expression values from the RNAi strain being normalised to this value. Means ± standard deviations are shown from triplicate cultures. Figure 4b was adapted from . Non-normalised GFP expression values are shown in Additional file : Figure S1.

Article Snippet: Plasmids P TEF1 -yEGFPCLN2PEST-pRS406, P SUC2 -yEGFPCLN2PEST-pRS406, and P SUC2 -GFPantisense-CYC1t-pRS413 will be made available from AddGene ( www.addgene.org/ ).

Techniques: Expressing, Control, Construct

Journal: Microbial Cell Factories

Article Title: Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae

doi: 10.1186/s12934-015-0223-7

Figure Lengend Snippet: S. cerevisiae strains used in this study

Article Snippet: Plasmids P TEF1 -yEGFPCLN2PEST-pRS406, P SUC2 -yEGFPCLN2PEST-pRS406, and P SUC2 -GFPantisense-CYC1t-pRS413 will be made available from AddGene ( www.addgene.org/ ).

Techniques: Expressing, Control

Journal: Microbial Cell Factories

Article Title: Dynamic regulation of gene expression using sucrose responsive promoters and RNA interference in Saccharomyces cerevisiae

doi: 10.1186/s12934-015-0223-7

Figure Lengend Snippet: Plasmids

Article Snippet: Plasmids P TEF1 -yEGFPCLN2PEST-pRS406, P SUC2 -yEGFPCLN2PEST-pRS406, and P SUC2 -GFPantisense-CYC1t-pRS413 will be made available from AddGene ( www.addgene.org/ ).

Techniques: Plasmid Preparation, Expressing, Selection, Marker, Construct

Journal: Journal of Visualized Experiments : JoVE

Article Title: Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae

doi: 10.3791/55836

Figure Lengend Snippet:

Article Snippet: Please click here to view a larger version of this figure. table ft1 table-wrap mode="anchored" t5 Plasmid Yeast selectable marker Notes Addgene plasmid number pGZ108 kanMX6 3xFLAG-MNase tagging, 33 aa linker 70231 pGZ109 HIS3MX6 3xFLAG-MNase tagging, 33 aa linker 70232 pGZ110 TRP1 3xFLAG-MNase tagging, 33 aa linker 70233 pGZ136 URA3 Expresses 3xFLAG-MNase-SV40 NLS under the control of the REB1 promoter 72273 pGZ173 kanMX6 MNase tagging, 8 aa linker 70234 Open in a separate window Table 1: Details of ChEC Plasmids.

Techniques: Plasmid Preparation, Marker, Control