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Metabolic Impacts of Using Nitrogen and Copper-Regulated Promoters to Regulate Gene Expression in Neurospora crassa.

Ouyang S, Beecher CN, Wang K, Larive CK, Borkovich KA - G3 (Bethesda) (2015)

Bottom Line: However, relatively few highly tunable promoters have been developed for this species.We determined that fragments corresponding to 1.5-kb fragments upstream of the tcu-1 and nit-6 open reading frames are needed for optimal repression and expression of GFP mRNA and protein. nit-6 was repressed using concentrations of glutamine from 2 to 20 mM and induced in medium containing 0.5-20 mM nitrate as the nitrogen source.Our findings demonstrate that nit-6 is a tunable promoter that joins tcu-1 as a choice for regulation of gene expression in N. crassa.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Pathology and Microbiology, University of California, Riverside, 900 University Avenue, Riverside, California 92521 College of Horticulture and Plant Protection, Yangzhou University, Yangzhou 225009, China.

No MeSH data available.


Related in: MedlinePlus

PCA results for the 1H NMR spectra of strain pnit-6_1.5 cultured on Gln (samples 1–6; blue) and nitrate (samples 7–12; red). (A) PCA scores. Although the samples within a treatment are not tightly clustered, clear segregation of the samples by treatments is observed in the scores plot. (B) PCA loadings. The loadings plot highlights the 1H NMR resonances that make the greatest contributions to sample variance in this data set. The loadings for PC1 are shown in blue, and those for PC2 are shown in green.
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fig5: PCA results for the 1H NMR spectra of strain pnit-6_1.5 cultured on Gln (samples 1–6; blue) and nitrate (samples 7–12; red). (A) PCA scores. Although the samples within a treatment are not tightly clustered, clear segregation of the samples by treatments is observed in the scores plot. (B) PCA loadings. The loadings plot highlights the 1H NMR resonances that make the greatest contributions to sample variance in this data set. The loadings for PC1 are shown in blue, and those for PC2 are shown in green.

Mentions: A clustering of the samples by treatment type is observed in the PCA score plots for the pnit-6_1.5(Gln) and pnit-6_1.5(nitrate) samples (Figure 5A). Together, PC1 and PC2 account for 57% of the variance in this sample set. Although the samples are not tightly grouped, there is clear segregation of the samples of pnit-6_1.5 grown on VM-Gln (samples 1–6) and grown on media containing nitrate (samples 7–12). The PCA loadings highlight the variables, in this case metabolite NMR resonances, responsible for the greatest variance for a particular principal component (e.g., PC1 or PC2). Analysis of the loadings plot (Figure 5B) confirms our observations that the levels of Gln, Glu, Ala, mannitol, and trehalose differ significantly in the NMR spectra measured for the two treatments. In addition, the PCA loadings highlight the resonances between 0.5 and 1.5 ppm due to amino acids (e.g., valine, leucine, isoleucine, and threonine) that we did not identify as important in our analysis of the NMR spectra. Although the intensity of the PCA loading for a particular variable is an indicator of its relative importance, it should be recognized that loadings reflect the variance over all samples and cannot necessarily be assigned to individual treatments. This is especially the case when, as in Figure 5A, the sample groupings separate along both PC1 and PC2. In addition, in interpreting these results it should be noted that the signs of the loadings are arbitrary and are not correlated to an increase or decrease in resonance intensity.


Metabolic Impacts of Using Nitrogen and Copper-Regulated Promoters to Regulate Gene Expression in Neurospora crassa.

Ouyang S, Beecher CN, Wang K, Larive CK, Borkovich KA - G3 (Bethesda) (2015)

PCA results for the 1H NMR spectra of strain pnit-6_1.5 cultured on Gln (samples 1–6; blue) and nitrate (samples 7–12; red). (A) PCA scores. Although the samples within a treatment are not tightly clustered, clear segregation of the samples by treatments is observed in the scores plot. (B) PCA loadings. The loadings plot highlights the 1H NMR resonances that make the greatest contributions to sample variance in this data set. The loadings for PC1 are shown in blue, and those for PC2 are shown in green.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4555226&req=5

fig5: PCA results for the 1H NMR spectra of strain pnit-6_1.5 cultured on Gln (samples 1–6; blue) and nitrate (samples 7–12; red). (A) PCA scores. Although the samples within a treatment are not tightly clustered, clear segregation of the samples by treatments is observed in the scores plot. (B) PCA loadings. The loadings plot highlights the 1H NMR resonances that make the greatest contributions to sample variance in this data set. The loadings for PC1 are shown in blue, and those for PC2 are shown in green.
Mentions: A clustering of the samples by treatment type is observed in the PCA score plots for the pnit-6_1.5(Gln) and pnit-6_1.5(nitrate) samples (Figure 5A). Together, PC1 and PC2 account for 57% of the variance in this sample set. Although the samples are not tightly grouped, there is clear segregation of the samples of pnit-6_1.5 grown on VM-Gln (samples 1–6) and grown on media containing nitrate (samples 7–12). The PCA loadings highlight the variables, in this case metabolite NMR resonances, responsible for the greatest variance for a particular principal component (e.g., PC1 or PC2). Analysis of the loadings plot (Figure 5B) confirms our observations that the levels of Gln, Glu, Ala, mannitol, and trehalose differ significantly in the NMR spectra measured for the two treatments. In addition, the PCA loadings highlight the resonances between 0.5 and 1.5 ppm due to amino acids (e.g., valine, leucine, isoleucine, and threonine) that we did not identify as important in our analysis of the NMR spectra. Although the intensity of the PCA loading for a particular variable is an indicator of its relative importance, it should be recognized that loadings reflect the variance over all samples and cannot necessarily be assigned to individual treatments. This is especially the case when, as in Figure 5A, the sample groupings separate along both PC1 and PC2. In addition, in interpreting these results it should be noted that the signs of the loadings are arbitrary and are not correlated to an increase or decrease in resonance intensity.

Bottom Line: However, relatively few highly tunable promoters have been developed for this species.We determined that fragments corresponding to 1.5-kb fragments upstream of the tcu-1 and nit-6 open reading frames are needed for optimal repression and expression of GFP mRNA and protein. nit-6 was repressed using concentrations of glutamine from 2 to 20 mM and induced in medium containing 0.5-20 mM nitrate as the nitrogen source.Our findings demonstrate that nit-6 is a tunable promoter that joins tcu-1 as a choice for regulation of gene expression in N. crassa.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Pathology and Microbiology, University of California, Riverside, 900 University Avenue, Riverside, California 92521 College of Horticulture and Plant Protection, Yangzhou University, Yangzhou 225009, China.

No MeSH data available.


Related in: MedlinePlus