Molecular cloning and heterologous expression of a 10-deacetylbaccatin III-10-O-acetyl transferase cDNA from Taxus x media.
A full-length cDNA encoding 10-deacetylbaccatin III-10-O-acetyl transferase (designated as TmDBAT), which catalyzes the acetylation of the C-10 hydroxyl group of the advanced metabolite 10-deacetylbaccatin III (10-DAB) to yield baccatin III, the immediate diterpenoid precursor of Taxol, was isolated from Taxus x media. Heterologous expression of TmDBAT in E. coli demonstrated that TmDBAT was a functional gene. Tissue expression pattern analysis revealed that TmDBAT expressed strongly in leaves, weak in stems and no expression could be detected in fruits, implying that TmDBAT was tissue-specific. Expression profiling analysis of TmDBAT under different elicitor treatments including silver nitrate, ammonium ceric sulphate and methyl jasmonate indicated that TmDBAT was an elicitor-responsive gene. Southern blot analysis suggested that TmDBAT belonged to a small multigene family.
Guo
Binhui
B
Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, School of Life Science and Technology, Shanghai Jiao Tong University, Shanghai, 200030, People's Republic of China.
Kai
Guoyin
G
Gong
Yifu
Y
Jin
Hongbin
H
Wang
Yechun
Y
Miao
Zhiqi
Z
Sun
Xiaofen
X
Tang
Kexuan
K
D000123:Acetyltransferases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D002678:Chimera; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D015870:Gene Expression; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D043823:Taxoids / Q000378:metabolism*; D020946:Taxus / Q000201:enzymology*
D018076:DNA, Complementary; D010940:Plant Proteins; D043823:Taxoids; C042213:10-deacetylbaccatine III; D000123:Acetyltransferases