Sub-cellular localization and overexpressing analysis of hydroxylase gene TcCYP725A22 of Taxus chinensis.

Weifang Liao; Chunhua FU; Zhiguo Liu; Lihong Miao; Longjiang YU

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Sub-cellular localization and overexpressing analysis of hydroxylase gene TcCYP725A22 of Taxus chinensis.

Author: Weifang LIAO ¹ ; Chunhua FU ² ; Zhiguo LIU ¹ ; Lihong MIAO ¹ ; Longjiang YU ²
Author Information

1. School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, Hubei, China.
2. School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, Hubei, China.
Publication Type:Journal Article
Keywords: Taxus chinensis; biosynthetic pathway; hydroxylase; overexpression; sub-cellular localization; taxol
MeSH: Biosynthetic Pathways; Mixed Function Oxygenases; Paclitaxel; Taxoids; Taxus
From: Chinese Journal of Biotechnology 2019;35(6):1109-1116
CountryChina
Language:Chinese
Abstract: The discovery of hydroxylases in the anticancer drug taxol biosynthesis pathway is a hotspot and difficulty in current research. In this study, a new hydroxylase gene TcCYP725A22 (GenBank accession number: MF448646.1) was used to construct a sub-cellular localization vector pCAMIBA1303-TcCYP725A22-EGFP to get the transient expression in onion epidermal cells. Laser confocal microscopy revealed that the protein encoded by this gene was localized in the cell membrane. Furthermore, the recombinant plant expression plasmid pBI121-TcCYP725A22 was constructed. After transient transformation to the Taxus chinensis mediated by Agrobacterium tumefaciens LBA4404, qRT-PCR and LC-MS were utilized to analyze the effects of TcCYP725A22 overexpression on the synthesis of taxol. The results showed that, in the TcCYP725A22 overexpressed cell line, expression levels of most defined hydroxylase genes for taxol biosynthesis were increased, and the yield of taxanes were also increased. It was concluded that the hydroxylase gene TcCYP725A22 is likely involved in the biosynthetic pathway of taxol.