Cloning, prokaryotic expression, and functional identification of a sesquiterpene synthase gene (AsSS4) from Aquilaria sinensis / 药学学报
Acta Pharmaceutica Sinica
;
(12): 1724-1729, 2014.
Article
Dans Chinois
| WPRIM
| ID: wpr-251829
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
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Polyisoprényl-phosphates
/
Sesquiterpènes
/
Protéines recombinantes
/
Cadres ouverts de lecture
/
Clonage moléculaire
/
ADN complémentaire
/
Alkyl et aryl transferases
/
Thymelaeaceae
/
Sesquiterpènes de type guaïane
/
Escherichia coli
langue:
Chinois
Texte intégral:
Acta Pharmaceutica Sinica
Année:
2014
Type:
Article
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