Journal: Synthetic and Systems Biotechnology

Article Title: Modifying the upstream open reading frames of cellulase gene enhances cellulase production in Penicillium oxalicum

doi: 10.1016/j.synbio.2025.12.016

Figure Lengend Snippet: Effects of functional uORFs in the 5′-UTR of the eg1 gene on gene expression in P . oxalicum grown on Avicel. (a) RT-qPCR analysis of gfp transcription. (b) RT-qPCR assay of eg1 transcription in OE- eg1 - muORF mutants. (c) mRNA stability of eg1 following 6 h dactinomycin pretreatment. (d) mRNA stability of eg1 following 2 h cycloheximide pretreatment. Data values represent mean ± standard deviation. Each experiment was performed with at least three biological replicates. ∗ p < 0.05 and ∗∗ p < 0.01 denote statistically significant differences compared to the control strain eg1 -5′-UTR- GFP or OE eg1 .

Article Snippet: The sample was resuspended in the cold polysome extraction buffer (200 mM tetramethylammonium hydroxide-HCl [pH 8.0], 30 mM potassium chloride, 15 mM magnesium chloride, 2 % [v/v] polyoxymethylene tridecyl ether, 100 mg/mL cycloheximide, 1 % [w/v] deoxycholic acid, 1.0 mg/mL dithiothreitol, 10 U/mL DNase I), and the mixture was kept on ice for 25 min. After centrifugation at 14,000 rpm for 30 min, the OD260 value of the supernatant was measured using a NanoDrop spectrophotometer (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Techniques: Functional Assay, Gene Expression, Quantitative RT-PCR, Standard Deviation, Control