期刊论文

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

中文

生物工程学报

1000-3061

2019

35

6

1109-1116

10.13345/j.cjb.190008

高等学校博士学科点专项科研基金（No.20120142130009）资助~~；

本校为第一机构

抗癌药物紫杉醇生物合成途径中羟化酶的发现是当今研究的热点和难点。文中利用前期分析得到的1 个新的中国红豆杉羟化酶基因TcCYP725A22 (GenBank 登录号：MF448646.1)，构建了亚细胞定位载体pCAMIBA1303-TcCYP725A22-EGFP，瞬时侵染洋葱表皮细胞，激光共聚焦显微镜观察结果发现该基因编码蛋白定位在细胞膜。进一步构建了植物表达载体pBI121-TcCYP725A22，经根癌农杆菌Agrobacterium tumefaciens LBA4404 介导转化到中国红豆杉细胞中过表达后，利用荧光定量PCR (qRT-PCR) 和液质联用(LC-MS) 分析TcCYP725A22 过表达对紫杉醇生物合成的影响。结果显示，瞬时转化pBI121-TcCYP725A22 的红豆杉细胞中...

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 T...