Molecular cloning and functional identification of a cDNA encoding 4-hydroxy-3-methylbut-2-enyl diphosphate reductase from.
10.1016/j.apsb.2016.12.002
- Author:
Qiqing CHENG
1
,
2
,
3
;
Yuru TONG
1
;
Zihao WANG
4
;
Ping SU
1
;
Wei GAO
4
;
Luqi HUANG
5
Author Information
1. School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China
2. National Resource Center for Chinese Materia Medica, Academy of Chinese Medical Sciences, Beijing 100700, China
3. State Key Laboratory of Quality Research in Chinese Medicines, Macau University of Science and Technology, Macau, China.
4. School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
5. National Resource Center for Chinese Materia Medica, Academy of Chinese Medical Sciences, Beijing 100700, China.
- Publication Type:Journal Article
- Keywords:
4-Hydroxy-3-methylbut-2-enyl diphosphate reductase;
Complementation;
Gene expression;
Tripterygium wilfordii;
Triptolide
- From:
Acta Pharmaceutica Sinica B
2017;7(2):208-214
- CountryChina
- Language:English
-
Abstract:
The 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR) is the last step key enzyme of the methylerythritol phosphate (MEP) pathway, synthesizing isopentenyl diphosphate and its allyl isomer dimethylallyl diphosphate, which is important for regulation of isoprenoid biosynthesis. Here the full-length cDNA of, designated(GenBank Accession No. KJ933412.1), was isolated fromfor the first time. TwHDR has an open reading frame (ORF) of 1386 bp encoding 461 amino acids. TwHDR exhibits high homology with HDRs of other plants, with an N-terminal conserved domain and three conserved cysteine residues.cDNA was cloned into an expression vector and transformed into anmutant. Since loss-of-functionmutant is lethal, the result showed that transformation ofcDNA rescued themutant. This complementation assay suggests that thecDNA encodes a functional HDR enzyme. The expression ofwas induced by methyl-jasmonate (MJ) insuspension cells. The expression ofreached the highest level after 1 h of MJ treatment. These results indicate that we have identified a functional TwHDR enzyme, which may play a pivotal role in the biosynthesis of diterpenoid triptolide in.
