Cloning, sequence analysis, and prokaryotic expression of GrGPPS gene in Gentiana rigescens / 中草药
Chinese Traditional and Herbal Drugs
;
(24): 2060-2068, 2014.
Artigo
em Chinês
| WPRIM
| ID: wpr-854465
ABSTRACT
Objective:
To obtain the indispensable key enzyme involved in the monoterpene biosynthesis, the geranyl diphosphate synthase gene GrGPPS was cloned from Gentiana rigescens, and its sequence analysis and prokaryotic expression were performed.Methods:
According to the GrGPPS gene sequence of transcriptome of triennial G. rigescens, a pair of specific primers was designed, and the full length of cDNA sequences was obtained by RT-PCR. Then TA cloning, sequencing and sequence analysing were performed. Prokaryotic expression vector pGEX-T-1-GrGPPS was constructed and transformed into Escherichia coli Rosetta for expression under 37°C and induced by 1 mmol/L IPTG.Results:
The GrGPPS cDNA had a length of 1107 bp coding for 369 amino acids. Sequence analysis showed that GrGPPS was the member of "short-chain prenyltransferases" super family. Results of phylogenic analysis showed that GrGPPS was at the same evolutionary branch with AmGPPS. The SDS-PAGE results displayed that the expressed proteins were consistent with the anticipated size.Conclusion:
The GrGPPS gene is cloned from G. rigescens, and the stable prokaryotic expression system of pGEX-T-1-GrGPPS is constructed. This work will provide a foundation for further purification, structure, and functional research of GrGPPS protein.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Chinese Traditional and Herbal Drugs
Ano de publicação:
2014
Tipo de documento:
Artigo
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