Cloning and functional analysis of squalene synthase gene from Dryopteris fragrans (L.) Schott.
Protein Expr Purif
; 155: 95-103, 2019 03.
Article
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| MEDLINE
| ID: mdl-30530061
ABSTRACT
Dryopteris fragrans (L.) Schott is a traditional herbal medicine containing medicinal sterols and triterpenoids. Squalene synthase (SQS) is the first crucial enzyme in the biosynthesis pathway of sterols and triterpenoids. The full-length cDNA named DfSQS1 was isolated by RACE. It was predicted that DfSQS1 contained an open reading frame (ORF) of 1239 bp coding 412 amino acid residues with molecular weight of 46.6â¯kDa. It had 18 potential phosphorylation sites, 1 potential N-glycosylation site and 2 transmembrane domains. In neighbor-joining (NJ) phylogenetic tree, DfSQS1 was away from branch of gymnosperms and angiosperms. One hydrophobic domain at the C-terminal of DfSQS1 was deleted to express soluble recombinant enzyme. The truncated DfSQS1 (tDfSQS1) was expressed in Escherichia coli BL21 (DE3). Then, tDfSQS1 was obtained and incubated with farnesyl diphosphate (FPP) to identify its enzymatic activity. The result demontrated that squalene, the product of enzyme catalyzed reaction, was detected by HPLC. Quantitative real-time PCR (qRT-PCR) analysis revealed that the transcription level of DfSQS1 in D. fragrans was the highest in roots, followed by leaves and rhizomes. This work is the first report on cloning, characteration and expression of SQS from D. fragrans. It will be helpful to understand the regulatory role of SQS on the biosynthesis of triterpenoids in the fern.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Proteínas de Plantas
/
Farnesil Difosfato Farnesil Transferasa
/
Dryopteris
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
Protein Expr Purif
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2019
Tipo del documento:
Article