[Cloning, prokaryotic expression, and functional identification of a sesquiterpene synthase gene (AsSS4) from Aquilaria sinensis].
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
Liang
L
Liang
Qing-Mei
QM
Guo
Zheng
Z
Zhang
Yan-Hong
YH
Xu
Xiao-Min
XM
Han
Juan
J
Liu
D019883:Alkyl and Aryl Transferases / Q000096:biosynthesis* / Q000235:genetics; D052176:Azulenes; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli; D000081222:Monocyclic Sesquiterpenes; D016366:Open Reading Frames; D011106:Polyisoprenyl Phosphates; D011994:Recombinant Proteins / Q000096:biosynthesis; D012717:Sesquiterpenes / Q000378:metabolism; D045789:Sesquiterpenes, Guaiane; D029645:Thymelaeaceae / Q000201:enzymology* / Q000235:genetics
D052176:Azulenes; D018076:DNA, Complementary; D000081222:Monocyclic Sesquiterpenes; D011106:Polyisoprenyl Phosphates; D011994:Recombinant Proteins; D012717:Sesquiterpenes; D045789:Sesquiterpenes, Guaiane; C445979:beta-elemene; C042686:humulene; C004808:farnesyl pyrophosphate; C019408:guaiene; D019883:Alkyl and Aryl Transferases; C108877:terpene synthase