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Transient Expression in Nicotiana Benthamiana Leaves for Triterpene Production at a Preparative Scale
Published on: August 16, 2018
[Cloning and protein expression analysis of monoterpene synthase gene TwMS in Tripterygium wilfordii]
Tian-Yuan Hu1, Ping Su2, Yi-Feng Zhang1,2
1Key Laboratory of Collateral Diseases, School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
Abstract:
In this study, we cloned a monoterpene synthases, TwMS from Tripterygium wilfordii suspension cells. TwMS gene contained a 1 797 bp open reading frame (ORF), encoding a polypeptide of 579 amino acids, which deduced isoelectric point (pI) was 6.10 and the calculated molecular weight was 69.75 kDa. Bioinformation analysis showed that the sequence of TwMS was consistent with the feature of monoterpene synthases. Differential expression analysis revealed that the relative expression level of TwMS increased significantly after being induced by methyl jasmonate (MeJA). The highest expression level occurred at 24 h. TwMS protein was successfully expressed in Escherichia coli BL21 (DE3), which laid the foundation for identifying the function of T. wilfordii monoterpene synthases.

