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Molecular cloning, expression and immunolocalization analysis of diphosphomevalonate decarboxylase involved in terpenoid biosynthesis from Euphorbia helioscopia L.
- Source :
-
Biotechnology & Biotechnological Equipment . Dec2017, Vol. 31 Issue 6, p1106-1115. 10p. - Publication Year :
- 2017
-
Abstract
- Euphorbia helioscopiaL. is an herbaceous species ofEuphorbia(Euphorbiaceae). As an ancient folk herbal medicine, the most effective medicinal component is terpenoid. Diphosphomevalonate decarboxylase (MDC) is the last rate-limiting enzyme of generating the isopentenyl pyrophosphate precursor of terpenoid in MVA pathway. The gene of MDC was cloned successfully fromE. helioscopia, and namedEhMDC(accession number: KP995936). The full length cDNA ofEhMDCwas 1653 bp, and it contained an open reading frame of 1245 bp, a 5′ untranslated region of 184 bp and a 3′ untranslated region of 224 bp. EhMDC contained 415 amino acids. Homologous sequence analysis showed that amino acid sequence of EhMDC had the highest identity of 88% withRicinus communisMDC. Phylogenetic analysis of EhMDC indicatedE. helioscopia,Hevea brasiliensis,R. communis, andJatropha carcaswhich all belonged to Euphorbiaceae were classified into one class. Real-time PCR assay demonstratedEhMDCwas constitutively expressed in roots, stems and leaves with a similar transcription level. Furthermore, in combination with immunoblot analysis and transmission electron microscopy immunogold labeling after anti-EhMDC antibody preparation, the EhMDC was found to be located in the cisternae of the endoplasmic reticulum, small vacuoles from endoplasmic reticulum and cytoplasm of laticifers. As a result, terpenoid biosynthesis site and accumulation ofE. helioscopialaticifers were speculated. [ABSTRACT FROM PUBLISHER]
- Subjects :
- *MOLECULAR cloning
*HERBAL medicine
*DECARBOXYLASES
*MESSENGER RNA
*GENE expression
Subjects
Details
- Language :
- English
- ISSN :
- 13102818
- Volume :
- 31
- Issue :
- 6
- Database :
- Academic Search Index
- Journal :
- Biotechnology & Biotechnological Equipment
- Publication Type :
- Academic Journal
- Accession number :
- 125566126
- Full Text :
- https://doi.org/10.1080/13102818.2017.1370677