Title Characterization of the effects of terminators and introns on recombinant gene expression in the basidiomycete Ceriporiopsis subvermispora
Author Dong Xuan Nguyen1,2, Emi Nishisaka1, Moriyuki Kawauchi1, Takehito Nakazawa1, Masahiro Sakamoto1, and Yoichi Honda1*
Address 1Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan, 2Biotechnology Center of Ho Chi Minh City, Ho Chi Minh City, Viet Nam
Bibliography Journal of Microbiology, 58(12),1037–1045, 2020,
DOI 10.1007/s12275-020-0213-2
Key Words Ceriporiopsis subvermispora, transient transformation, gpd gene, polyadenylation signals, intron functions
Abstract 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.