Journal: bioRxiv

Article Title: Ustilago maydis serves as a novel production host for the synthesis of plant and fungal sesquiterpenoids

doi: 10.1101/2020.05.04.076794

Figure Lengend Snippet: (A) Quantification of Gfp expression using fluorimeter measurements. Relative fluorescence units are given relative to the optical density (OD 600 ). At least three independent biological experiments (n=3) were performed with three technical replicates per strain. Error bars indicate standard error of the mean (SEM). Statistical significance was calculated using the unpaired two-tailed t test and p -values were indicated above. Note, that the AaCrtB Myc expressing strains carried a deletion of car2 . (B) Microscopic analysis showing DIC images of fixed cells on the left (size bar, 10 μm). Corresponding staining of DNA with Hoechst 33342 (middle panel; N, nucleus; inverted image) and green fluorescence (Gfp) on the right (yellow and red arrowheads indicate nuclei and peroxisomes, respectively).

Article Snippet: The excitation of fluorescently labelled proteins was carried out using an HXP metal halide lamp (LEj, Jena, Germany) in combination with a filter set for green fluorescent protein (ET470/40BP, ET495LP and ET525/50BP) and Hoechst 33342 dye (HC387/11BP, BS409LP and HC 447/ 60BP; AHF Analysentechnik, Tübingen, Germany).

Techniques: Expressing, Fluorescence, Two Tailed Test, Staining

Journal: bioRxiv

Article Title: Ustilago maydis serves as a novel production host for the synthesis of plant and fungal sesquiterpenoids

doi: 10.1101/2020.05.04.076794

Figure Lengend Snippet: (A) Quantification of Gfp expression using fluorimeter measurements. Relative fluorescence units are given relative to the optical density (OD 600 ). At least three independent biological experiments (n=3) were performed with three technical replicates per strain. Error bars indicate standard error of the mean (SEM). Statistical significance was calculated using the unpaired two-tailed t test and p -values were indicated above. Note, that the AaCrtB Myc expressing strains carried a deletion of car2 . (B) Microscopic analysis showing DIC images of fixed cells on the left (size bar, 10μm). Corresponding staining of nuclear DNA with Hoechst 33342 (middle panel; N, nucleus; inverted image) and green fluorescence (Gfp) on the right (yellow arrowheads indicate nuclei). (C) Cell pellets and lycopene concentrations (orange bars) of strains given at the bottom. Three independent biological experiments (n=3) were carried out. Error bars indicate standard deviation of the mean (SD). Statistical significance was calculated using the unpaired two-tailed t test and p -values were indicated above. Note, that the AaCrtB Myc expressing strains carried a deletion of car2 . (D) GC-MS chromatogram of (+)-valencene from CnVS expressing strain and the corresponding standard diluted in n-dodecane samples of the negative control. (E) Fragmentation pattern of peaks at 7.3 min (shown in D). (F) Concentration of (+)-valencene produced in the culture, determined by GC-FID according to commercial reference compound (Supplementary Figure S3B). Three independent biological experiments (n=3) were carried out. Error bars indicate standard deviation of the mean (SD). Statistical significance was calculated using the unpaired two-tailed t test and p -values were indicated above. Note, that the AaCrtB Myc expressing strains carried a deletion of car2 .

Article Snippet: The excitation of fluorescently labelled proteins was carried out using an HXP metal halide lamp (LEj, Jena, Germany) in combination with a filter set for green fluorescent protein (ET470/40BP, ET495LP and ET525/50BP) and Hoechst 33342 dye (HC387/11BP, BS409LP and HC 447/ 60BP; AHF Analysentechnik, Tübingen, Germany).

Techniques: Expressing, Fluorescence, Two Tailed Test, Staining, Standard Deviation, Gas Chromatography-Mass Spectrometry, Negative Control, Concentration Assay, Produced

Journal: bioRxiv

Article Title: Ustilago maydis serves as a novel production host for the synthesis of plant and fungal sesquiterpenoids

doi: 10.1101/2020.05.04.076794

Figure Lengend Snippet: (A) Quantification of Gfp expression using fluorimeter measurements. Relative fluorescence units (RFU) are given relative to the optical density (OD 600 ). At least three independent biological experiments (n=3) were performed with three technical replicates per strain. Error bars indicate standard error of the mean (SEM). Statistical significance was calculated using the unpaired two-tailed t test and p -values were indicated above. Note, that the AaCrtB Myc expressing strains carried a deletion of car2 . (B) Microscopic analysis showing DIC images of fixed cells on the left (size bar, 10 μm). Corresponding staining of DNA Hoechst 33342 (middle panel; N, nucleus, inverted image) and green fluorescence (Gfp) on the right (yellow arrowheads indicate nuclei). (C) Cell pellets and lycopene concentrations (orange bars) of strains given at the bottom. Three independent biological experiments (n=3) were carried out. Error bars indicate standard deviation of the mean (SD). Statistical significance was calculated using the unpaired two-tailed t test and p-values were indicated above. Note, that the AaCrtB Myc expressing strains carried a deletion of car2 . (D) GC-MS chromatogram of α-cuprenene from Cop6 expressing strains. (E) Fragmentation pattern of peaks at 7.3 min given in (D). (F) Estimated concentration of α-cuprenene determined via GC-FID using the reference compound β-caryophyllene (Supplementary Figure S4B). Three independent biological experiments (n=3) were carried out. Error bars indicate standard deviation of the mean (SD). Statistical significance was calculated using the unpaired two-tailed t test and p -values were indicated above. Note, that the AaCrtB Myc expressing strains carried a deletion of car2 .

Article Snippet: The excitation of fluorescently labelled proteins was carried out using an HXP metal halide lamp (LEj, Jena, Germany) in combination with a filter set for green fluorescent protein (ET470/40BP, ET495LP and ET525/50BP) and Hoechst 33342 dye (HC387/11BP, BS409LP and HC 447/ 60BP; AHF Analysentechnik, Tübingen, Germany).

Techniques: Expressing, Fluorescence, Two Tailed Test, Staining, Standard Deviation, Gas Chromatography-Mass Spectrometry, Concentration Assay