Showing papers on "Upstream activating sequence published in 2022"
TL;DR: A random mutagenesis library consisting of 330 different variants based on the UAS of the TDH3 promoter with an ∼37-fold dynamic range by error-prone polymerase chain reaction (PCR) is created and obtained strong intensity mutant UAS, which was ∼12-fold greater than the wild-type UASTDH3.
Abstract: A wide dynamic range of promoters is necessary for fine-tuning transcription levels. However, weak intensity and narrow dynamic range limit transcriptional regulation via constitutive promoters. The upstream activation sequence (UAS) located upstream of the core promoter is a crucial region that could obviously enhance promoter strength. Herein, we created a random mutagenesis library consisting of 330 different variants based on the UAS of the TDH3 promoter with an ∼37-fold dynamic range by error-prone polymerase chain reaction (PCR) and obtained strong intensity mutant UAS, which was ∼12-fold greater than the wild-type UASTDH3. Analysis of the mutant library revealed 15 strength-enhancing sites and their corresponding bases of the UASTDH3 regions, which provided the impetus for a synthetic library. The resulting 32 768 mutant UAS library was constructed by permutation and combination of the bases of the 15 enhancing sites. To characterize the library, a strength prediction model was built by correlating DNA structural features and UAS strength, which provided a model between UAS sequence and intensity. Following characterization, the UAS library was applied to precisely regulate gene expression in the production of β-carotene, proving that the UAS library would be a useful tool for gene tuning in metabolic engineering. In summary, we designed, constructed, and characterized a UAS library that facilitated precise tuning of transcription levels of target proteins.
4 citations
TL;DR: The xylP promoter from Penicillium chrysogenum was demonstrated to function in various mold species including Aspergillus fumigatus as mentioned in this paper , and structural and functional elements of pxylP were identified.
3 citations
TL;DR: In this paper , the authors constructed an hsp18-EGFP transcriptional fusion in an E. coli-Mycobacterium shuttle vector, and the integration of the construct was confirmed by Southern hybridization.