Identifying CYP Candidate Genes in T. wilfordii

CYPs were obtained from the PFAM database and annotated as cytochrome P450 (PF00067). A heatmap of gene expression was generated using MultiExperiment Viewer (MeV, version 4.9.0) and Java 8 software. The RPKM reads of the root periderm, root phloem, root xylem, stem vascular bundle, stem periderm, and leaf originated from previous tissue transcriptomes (NCBI SRA number SRP199495, Supplementary Fig. 20)^{16 (link)}, and the data were normalized and processed by hierarchical cluster analysis with Pearson correlation. CYPs with gene expression profiles similar to those of TwTPS7(v2), TwTPS27(v2), and CYP728B70 were selected as candidate CYPs for further analysis. The candidate CYPs were amplified using 2×Phusion High-Fidelity PCR Master Mix (New England Biolabs, USA) and cDNA of T. wilfordii root as the template, which was consistent with the transcriptome after complete sequencing (Supplementary Data 5).

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Zhang Y., Gao J., Ma L., Tu L., Hu T., Wu X., Su P., Zhao Y., Liu Y., Li D., Zhou J., Yin Y., Tong Y., Zhao H., Lu Y., Wang J., Gao W, & Huang L. (2023). Tandemly duplicated CYP82Ds catalyze 14-hydroxylation in triptolide biosynthesis and precursor production in Saccharomyces cerevisiae. Nature Communications, 14, 875.

Publication 2023

2×Phusion High-Fidelity PCR Master Mix

Cdna Cyps Gene expression Leaf Phloem Root Stem Tissue Transcriptome Vascular Xylem

Corresponding Organization :

Other organizations : Chinese Academy of Medical Sciences & Peking Union Medical College, Capital Medical University, Zhejiang University, Hangzhou Normal University, Zhejiang University of Technology, Beijing Shijitan Hospital

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Variable analysis

independent variables

Tissue types (root periderm, root phloem, root xylem, stem vascular bundle, stem periderm, and leaf)

dependent variables

RPKM (Reads Per Kilobase of transcript per Million mapped reads) of CYP (cytochrome P450) genes

control variables

CDNA of T. wilfordii root as the template for CYP gene amplification, which was consistent with the transcriptome after complete sequencing

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