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KMID : 0880220200580121037
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Journal of Microbiology
2020 Volume.58 No. 12 p.1037 ~ p.1045
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Characterization of the effects of terminators and introns on recombinant gene expression in the basidiomycete Ceriporiopsis subvermispora
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Nguyen Dong Xuan
Nishisaka Emi
Kawauchi Moriyuki
Nakazawa Takehito
Sakamoto Masahiro
Honda Yoichi
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Abstract
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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 Csβ1-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.
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KEYWORD
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Ceriporiopsis subvermispora, transient transformation, gpd gene, polyadenylation signals, intron functions
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