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Molecules 2018, 23(2), 269; https://doi.org/10.3390/molecules23020269

Cloning, Expression Analysis and Functional Characterization of Squalene Synthase (SQS) from Tripterygium wilfordii

1
Research & Development Center of Biorational Pesticides, Northwest A & F University, Yangling 712100, China
2
Biopesticide Technology and Engineering Center of Shaanxi Province, Yangling 712100, China
*
Authors to whom correspondence should be addressed.
Received: 4 January 2018 / Revised: 24 January 2018 / Accepted: 25 January 2018 / Published: 29 January 2018
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Abstract

Celastrol is an active triterpenoid compound derived from Tripterygium wilfordii which is well-known as a traditional Chinese medicinal plant. Squalene synthase has a vital role in condensing two molecules of farnesyl diphosphate to form squalene, a key precursor of triterpenoid biosynthesis. In the present study, T. wilfordii squalene synthase (TwSQS) was cloned followed by prokaryotic expression and functional verification. The open reading frame cDNA of TwSQS was 1242 bp encoding 413 amino acids. Bioinformatic and phylogenetic analysis showed that TwSQS had high homology with other plant SQSs. To obtain soluble protein, the truncated TwSQS without the last 28 amino acids of the carboxy terminus was inductively expressed in Escherichia coli Transetta (DE3). The purified protein was detected by SDS-PAGE and Western blot analysis. Squalene was detected in the product of in vitro reactions by gas chromatograph-mass spectrometry, which meant that TwSQS did have catalytic activity. Organ-specific and inducible expression levels of TwSQS were detected by quantitative real-time PCR. The results indicated that TwSQS was highly expressed in roots, followed by the stems and leaves, and was significantly up-regulated upon MeJA treatment. The identification of TwSQS is important for further studies of celastrol biosynthesis in T. wilfordii. View Full-Text
Keywords: Tripterygium wilfordii; squalene synthase; celastrol; prokaryotic expression; functional characterization Tripterygium wilfordii; squalene synthase; celastrol; prokaryotic expression; functional characterization
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Zhang, B.; Liu, Y.; Chen, M.; Feng, J.; Ma, Z.; Zhang, X.; Zhu, C. Cloning, Expression Analysis and Functional Characterization of Squalene Synthase (SQS) from Tripterygium wilfordii. Molecules 2018, 23, 269.

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