Molecular cloning, characterization and heterologous expression in Saccharomyces cerevisiae of a mevalonate diphosphate decarboxylase cDNA from Ginkgo biloba

Abstract

Mevalonate diphosphate decarboxylase (MVD; EC4.1.1.33) catalyses the conversion of mevalonate diphosphate to isopentenyl diphosphate (IPP), which is a key skeleton for numerous functionally important secondary metabolites. A full‐length cDNA of MVD was isolated from Ginkgo biloba (designated as GbMVD) using rapid amplification of cDNA ends (RACE). GbMVD was 1958 bp with poly(A) tailing and contained a 1290‐bp open reading frame encoding a 430‐amino acid protein. The deduced GbMVD protein had a deduced molecular weight of about 48 kDa and isoelectric point of 5.83, and it showed high similarity to MVDs from other species. The 3D‐structure modelling showed that the 3D model of GbMVD had a similar P loop as that of yeast. Phylogenetic tree analysis revealed that GbMVD shared the same ancestor with MVDs from other species, and it had a closer relationship with those from plant species. Southern blot analysis indicated that GbMVD might belong to a small multigene family. Transcript accumulation analysis revealed that GbMVD was transcribed in root, stem and leaf tissues. Functional complementation of GbMVD in an MVD‐deficient Saccharomyces cerevisiae strain MN19‐34 (erg19ts) confirmed that GbMVD mediated the conversion of mevalonate diphosphate to IPP and that it is a functional gene.

Characterization and Transcriptional Profiling of Ginkgo biloba Mevalonate Diphosphate Decarboxylase Gene (GbMVD) Promoter Towards Light and Exogenous Hormone Treatments

“…GbMVD Transcript Levels by Ginkgo Organs Pang et al (2006) showed that GbMVD was expressed in the roots, stems, and leaves. We extended the analysis of the organ expression pattern of GbMVD to additional organs of ginkgo via qRT-PCR.…”

“…Ginkgo genome walker libraries were constructed using the Genome Walker Universal Kit User (Clontech, USA). To clone the promoter region of GbMVD, two rounds of PCR were performed using gene-specific primers (GbMVDP1 and GbMVDP2) that were designed according to the sequence of GbMVD cDNA (AY757921; Pang et al, 2006) and the adapter primers (AP1 and AP2) provided in the kit. The primers are listed in Table 1.…”

Molecular cloning, expression and immunolocalization analysis of diphosphomevalonate decarboxylase involved in terpenoid biosynthesis from Euphorbia helioscopia L.

“…EhMDC in roots was more abundant than that in stems and leaves (Figure 4). Pang et al (2006) tested the GbMVD expression in Ginkgo biloba using RT-PCR. They found that GbMVD was transcribed in root, stem and leaf, and there was no distinct difference in their transcription levels [28].…”

“…Pang et al (2006) tested the GbMVD expression in Ginkgo biloba using RT-PCR. They found that GbMVD was transcribed in root, stem and leaf, and there was no distinct difference in their transcription levels [28]. To assess the organ-specific and environmentally responsive characteristics of GbMVD expression, Liao et al (2016) fused GbMVDpro to GUS reporter gene.…”

Biosynthesis pathways of ginkgolides

“…A full-length cDNA of MVD from G. biloba (GbMVD) was isolated and characterized. [23] The full-length cDNA of GbMVD was 1,958 bp with a poly (A) tailing, containing an ORF of 1,290 bp encoding 430 amino acids. There was a 5’-UTR of 291 bp upstream from the start codon with G as putative transcription start, and the coding region was followed by 3’-UTR that was 345 bp-long downstream from the stop codon.…”