To explore whether the farnesyl pyrophosphate synthase gene(DnFPPS)in Dendrobium nobile is in-volved in the metabolic processes such as sesquiterpene alkaloid and terpene synthesis,in this study,the full-length cDNA sequence of FPPS gene of Dendrobium nobile according to the reanalysis of transcriptome data published in NCBI for Dendrobium nobile was cloned;DnFPPS+PGEX-6t-1 prokaryotic expression vector was constructed and expressed in E.coli.A total of 1 044 bp nucleotides of FPPS gene of Dendrobium nobile were obtained.The NCBI blastx showed that the protein encoded by DnFPPS gene was the highest homology with FPPS1 from Dendrobium huoshanense,and the consistency of amino acids was 98％.Five typical conserved domains of isopentenyl trans-ferase gene existed in DnFPPS gene.Close genetic relationship was found between DnFPPS protein to DhFPPS1 by phylogenetic tree analysis.The predicted protein structure was 73.16％homologous with the crystal structure of FPP/GFPP synthetase in Artemisia spiciformis.RT-qPCR result revealed that DnFPPS gene expression was posi-tively correlated with dendrobine accumulation.SDS-PAGE electrophoresis observed about 72 kD bands,which was consistent with the predicted protein molecular weight.This study laid a key foundation for the functional study of DnFPPS gene and provided an important theoretical reference for elucidating the biosynthesis pathway of sesquiterpenoids and sesquiterpenoid alkaloids in Dendrobium nobile.
维普
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