2025-10-23 北海道大学
本研究の概要
<関連情報>
- https://www.hokudai.ac.jp/news/2025/10/post-2096.html
- https://www.hokudai.ac.jp/news/pdf/251023_pr2.pdf
- https://bmcplantbiol.biomedcentral.com/articles/10.1186/s12870-025-07411-y
緑藻スジアオノリにおける抗生物質選択マーカー遺伝子の安定発現とCRISPRを用いたノックインシステムの開発 Stable transgene expression and CRISPR-mediated knock-in system of a bacteria-derived antibiotic selection gene in the green alga Ulva prolifera
Zheng Qin,Whelver Surnido,Hiroyuki Mizuta & Toshiki Uji
BMC Plant Biology Published:06 October 2025
DOI:https://doi.org/10.1186/s12870-025-07411-y
Abstract
Ulva prolifera is a fast-growing green seaweed that has garnered considerable interest in both fundamental and applied research. Here, we established a molecular tool by employing a selectable marker gene that allowed the isolation of U. prolifera cells integrating exogenous DNA. We developed a modular plasmid for expressing exogenous genes in U. prolifera based on the bacterial antibiotic-resistance marker, aminoglycoside phosphotransferase gene (aph7”). Integration of aph7” in macroalgae can generate transformants resistant to hygromycin B. In addition, we characterized the promoter region of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase gene (pUpRbcS) to drive the expression of aph7”. The transcripts were consistently confirmed from antibiotic-selected transformants, stably retaining the exogenous gene in the succeeding generations. Subsequently, a CRISPR-based knock-in system was established, facilitating the integration of aph7” cassette in the endogenous selection gene encoding for adenine phosphoribosyltransferase (UpAPT). APT gene can serve as an endogenous marker in algae that exhibits a lethal phenotype under cultivation with 2-fluoroadenine. The resulting knock-in mutants could resist the co-selection of the antibiotic hygromycin B and 2-fluoroadenine. Our results advance U. prolifera as a genetic platform, enabling functional research to elucidate Ulva biology, and to bring forth biotechnological utilization of algal resources.
