1. [Cloning, prokaryotic expression, and functional identification of a sesquiterpene synthase gene (AsSS4) from Aquilaria sinensis].
- Author
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Liang L, Guo QM, Zhang Z, Xu YH, Han XM, and Liu J
- Subjects
- Alkyl and Aryl Transferases genetics, Azulenes, Cloning, Molecular, DNA, Complementary, Escherichia coli, Monocyclic Sesquiterpenes, Open Reading Frames, Polyisoprenyl Phosphates, Recombinant Proteins biosynthesis, Sesquiterpenes metabolism, Sesquiterpenes, Guaiane, Thymelaeaceae genetics, Alkyl and Aryl Transferases biosynthesis, Thymelaeaceae enzymology
- Abstract
A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis
- Published
- 2014