The protein sequences of OsS5H and mutated OsS5H were uploaded to the publicly accessible Phyre2 website (http://www.sbg.bio.ic.ac.uk/phyre2 ; Kelley et al., 2015 (link)) for modelling. Then protein models were further edited with ChimeraX software (Pettersen et al., 2021 (link)).
To perform Y2H assay, CDS of OsS5H2, OsS5H3, and OsS5H4 was constructed into vector pGBKT7 (BD), and truncated C‐terminus of OsUGT74H4 was cloned into pGADT7 (AD). Yeast strain AH109 was used for transformation, and the detailed experimental process was described in a report by Liu, Cai et al. (2020 (link)).
To perform Y2H assay, CDS of OsS5H2, OsS5H3, and OsS5H4 was constructed into vector pGBKT7 (BD), and truncated C‐terminus of OsUGT74H4 was cloned into pGADT7 (AD). Yeast strain AH109 was used for transformation, and the detailed experimental process was described in a report by Liu, Cai et al. (2020 (link)).
