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.