Cloning and expression analysis of GGPPS gene from Panax notoginseng.
- Author:
Dan-dan MIN
;
Mei-qiong TANG
;
Gang LI
;
Xiao-sheng QU
;
Jian-hua MIAO
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Computational Biology;
Geranylgeranyl-Diphosphate Geranylgeranyltransferase;
genetics;
Panax notoginseng;
genetics;
Real-Time Polymerase Chain Reaction
- From:
China Journal of Chinese Materia Medica
2015;40(11):2090-2095
- CountryChina
- Language:Chinese
-
Abstract:
According to the transcriptome dataset of Panax notoginseng, the key geranylgeranyl pyrophosphate synthase gene (GGPPS) in terpenoid backbone biosynthesis was selected to be cloned. Using specific primer pairs combining with RACE (rapid amplification of cDNA ends) technique, the full-length cDNA sequence with 1 203 bp, which containing a 1 035 bp open reading frame, was cloned and named as PnGGPPS. The corresponding full-length DNA sequence contained 2 370 bp, consisted of 1 intron and 2 exons. The deduced protein PnGGPPS contained 344 amino acids and shared more than 73% identity with GGPPS from Ricinus communis and Salvia miltiorrhiza. PnGGPPS also had specific Aspartic acid enrichment regions and other conserved domains, which belonged to the Isoprenoid-Biosyn-C1 superfamily. The quantitative real-time PCR showed that PnGGPPS expressed in different tissues of 1, 2, 3 years old root, stem, leaf and 3 years old flower, and the expression level in 3 years old leaf was significant higher than that in other organs, which suggested that it might not only be involved in the regulation of the growth and development, but also be associated with the biosynthesis of chlorophyll and carotenoids, the development of chloroplast, the shade habit and the quality formation of P. notoginseng.
