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.
Artigo
em Chinês
| 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
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Fosfatos de Poli-Isoprenil
/
Sesquiterpenos
/
Proteínas Recombinantes
/
Fases de Leitura Aberta
/
Clonagem Molecular
/
DNA Complementar
/
Alquil e Aril Transferases
/
Thymelaeaceae
/
Sesquiterpenos de Guaiano
/
Escherichia coli
Idioma:
Chinês
Revista:
Acta Pharmaceutica Sinica
Ano de publicação:
2014
Tipo de documento:
Artigo
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