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Characterization of the effects of terminators and introns on recombinant gene expression in the basidiomycete Ceriporiopsis subvermispora
Dong Xuan Nguyen , Emi Nishisaka , Moriyuki Kawauchi , Takehito Nakazawa , Masahiro Sakamoto , Yoichi Honda
J. Microbiol. 2020;58(12):1037-1045.   Published online September 30, 2020
DOI: https://doi.org/10.1007/s12275-020-0213-2
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AbstractAbstract
Terminators and introns are vital regulators of gene expression in many eukaryotes; however, the functional importance of these elements for controlling gene expression in Agaricomycetes remains unclear. In this study, the effects of Ceriporiopsis subvermispora terminators and introns on the expression of a recombinant hygromycin B phosphotransferase gene (hph) were characterized. Using a transient transformation system, we proved that a highly active terminator (e.g., the gpd terminator) is required for the efficient expression of the hph gene. Mutational analyses of the C. subvermispora gpd terminator revealed that hph expression was dictated by an A-rich region, which included a putative positioning element, and polyadenylation sites. In contrast, our results indicated that introns are not required for the expression of hph directed by the CsĪ²1-tub and Csgpd promoters in C. subvermispora. This study provides insights into the functions and cis-element requirements of transcriptional terminators in Agaricomycetes, which may be relevant for designing recombinant genes for this important fungal class.

Citations

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  • Development of a 2A peptide-based multigene expression system and its application for enhanced production of ganoderic acids in Ganoderma lucidum
    Qiong Wang, Hong-Jun Liu, Yan Xu, Zi-Xu Wang, Bin Sun, Jun-Wei Xu
    Journal of Biotechnology.2024; 393: 109.     CrossRef
  • CRISPR/Cas9 using a transient transformation system in Ceriporiopsis subvermispora
    Takehito Nakazawa, Chikako Inoue, Dong Xuan Nguyen, Moriyuki Kawauchi, Masahiro Sakamoto, Yoichi Honda
    Applied Microbiology and Biotechnology.2022; 106(17): 5575.     CrossRef
  • A promoter assay system using gene targeting in agaricomycetes Pleurotus ostreatus and Coprinopsis cinerea
    Dong Xuan Nguyen, Takehito Nakazawa, Genki Myo, Chikako Inoue, Moriyuki Kawauchi, Masahiro Sakamoto, Yoichi Honda
    Journal of Microbiological Methods.2020; 179: 106053.     CrossRef

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