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Cloning and expression analysis of two dehydrodolichyl diphosphate synthase genes from Tripterygium wilfordii
Lin-Hui Gao1, Ping Su2, Yi-Feng Zhang1, Li-Chan Tu3, Yu-Jun Zhao2, Tian-Yuan Hu3, Jia-Wei Zhou3, Bao-Wei Ma3, Wei Gao4, Lu-Qi Huang2
1 School of Traditional Chinese Medicine, Capital Medical University; State Key Laboratory of Breeding Base Dao-di Herbs, National Resource Center for Chinese Materia Medical, China Academy of Chinese Medical Sciences, Beijing, PR China
2 State Key Laboratory of Breeding Base Dao-di Herbs, National Resource Center for Chinese Materia Medical, China Academy of Chinese Medical Sciences, Beijing, PR China
3 School of Traditional Chinese Medicine, Capital Medical University, Beijing, PR China
4 School of Traditional Chinese Medicine, Capital Medical University; Beijing Key Laboratory of Traditional Chinese Medicine Collateral Disease Theory Research, Capital Medical University, Beijing, PR China
Correspondence Address:
Lu-Qi Huang
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medical, China Academy of Chinese Medical Sciences, Beijing 100700
PR China
Wei Gao
School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069
PR China
 Source of Support: None, Conflict of Interest: None  | 12 |
DOI: 10.4103/wjtcm.wjtcm_4_18
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Objective: To clone and investigate two dehydrodolichyl diphosphate synthase genes of Tripterygium wilfordii by bioinformatics and tissue expression analysis. Materials and Methods: According to the T. wifordii transcriptome database, specific primers were designed to clone the TwDHDDS1 and TwDHDDS2 genes via PCR. Based on the cloned sequences, protein structure prediction, multiple sequence alignment and phylogenetic tree construction were performed. The expression levels of the genes in different tissues of T. wilfordii were measured by real-time quantitative PCR. Results: The TwDHDDS1 gene encompassed a 873 bp open reading frame (ORF) and encoded a protein of 290 amino acids. The calculated molecular weight of the translated protein was about 33.46 kDa, and the theoretical isoelectric point (pI) was 8.67. The TwDHDDS2 encompassed a 768 bp ORF, encoding a protein of 255 amino acids with a calculated molecular weight of about 21.19 kDa, and a theoretical isoelectric point (pI) of 7.72. Plant tissue expression analysis indicated that TwDHDDS1 and TwDHDDS2 both have relatively ubiquitous expression in all sampled organ tissues, but showed the highest transcription levels in the stems. Conclusions: The results of this study provide a basis for further functional studies of TwDHDDS1 and TwDHDDS2. Most importantly, these genes are promising genetic targets for the regulation of the biosynthetic pathways of important bioactive terpenoids such as triptolide.
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