Novel assays to monitor gene expression and protein-protein interactions in rice using the bioluminescent protein, NanoLuc
| Autor: | Koji Yamaguchi, Mana Ogawa, Zenpei Shimatani, Hiroyuki Tsuji, Hiromi Saitoh, Tsutomu Kawasaki, Yoichi Ikeda, Hiroko Akashi, Ken-ichiro Taoka, Rie Terada |
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| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
Original Paper 0303 health sciences Oryza sativa food and beverages Plant Science Biology 01 natural sciences Cell biology Complementation 03 medical and health sciences chemistry.chemical_compound chemistry Gene knockin Gene expression Bioluminescence Luciferase Florigen Agronomy and Crop Science Gene 030304 developmental biology 010606 plant biology & botany Biotechnology |
| Zdroj: | Plant Biotechnol (Tokyo) |
| ISSN: | 1347-6114 1342-4580 |
| DOI: | 10.5511/plantbiotechnology.20.1209a |
| Popis: | Luciferases have been widely utilized as sensitive reporters to monitor gene expression and protein-protein interactions. Compared to firefly luciferase (Fluc), a recently developed luciferase, Nanoluciferase (NanoLuc or Nluc), has several superior properties such as a smaller size and stronger luminescence activity. We compared the reporter properties of Nluc and Fluc in rice (Oryza sativa). In both plant-based two-hybrid and split luc complementation (SLC) assays, Nluc activity was detected with higher sensitivity and specificity than that with Fluc. To apply Nluc to research involving the photoperiodic regulation of flowering, we made a knock-in rice plant in which the Nluc coding region was inserted in-frame with the OsMADS15 gene, a target of the rice florigen Hd3a. Strong Nluc activity in response to Hd3a, and in response to change in day length, was detected in rice protoplasts and in a single shoot apical meristem, respectively. Our results indicate that Nluc assay systems will be powerful tools to monitor gene expression and protein-protein interaction in plant research. |
| Databáze: | OpenAIRE |
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