Molecular cloning of farnesyl pyrophosphate synthase from Alisma orientale (Sam.) Juzep. and its distribution pattern and bioinformatics analysis.
- Author:
Wei GU
1
;
Qi-nan WU
;
Jian-guo CHAO
;
Bei-li XI
;
Lin LI
;
Xiu-yuan SHEN
Author Information
1. College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210046, China. guwei9926@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Alisma;
enzymology;
genetics;
Amino Acid Sequence;
Base Sequence;
Cloning, Molecular;
Computational Biology;
Conserved Sequence;
DNA, Complementary;
genetics;
DNA, Plant;
genetics;
Gene Amplification;
Geranyltranstransferase;
genetics;
isolation & purification;
metabolism;
Molecular Sequence Data;
Phylogeny;
Plant Leaves;
enzymology;
genetics;
Plant Roots;
enzymology;
genetics;
Plants, Medicinal;
enzymology;
genetics;
RNA, Messenger;
metabolism;
Real-Time Polymerase Chain Reaction;
methods
- From:
Acta Pharmaceutica Sinica
2011;46(5):605-612
- CountryChina
- Language:Chinese
-
Abstract:
Triterpenes, which have large application potential in the treatment of cancer, are the main active components of genuine medicinal material Alisma orientale (Sam.) Juzep. Farnesyl pyrophosphate synthase (FPPS) is one of the important rate-limiting enzymes in the synthetic pathway of triterpenes. In this study the FPPS full length cDNA of the A. orientale, was cloned via homology-based cloning approach and rapid amplification of cDNA ends (RACE). The full length of the FPPS cDNA was 1 531 bp (accession no. HQ724508), which contained a full 1 032 bp ORF that encoded 343 amino acids. The deduced protein sequence exhibited five conserved motifs, two of which is riched of Asp (DDXXD). The result of real-time quantitative PCR (QRT-PCR) showed that FPPS gene was expressed in different organs of A. orientale. The expression increased from October to the first ten-day period of December, and then decreased. The FPPS gene expression was higher in leaves but lower in leafstalk, tuber and root. HPLC analysis of active components 23-acetyl-alismol B of A. orientale. during different periods indicated that its change trend should be consistent with FPPS gene expression. It can be primarily deduced that FPPS gene should be an important control point in the synthetic pathway of Alisma terpenes. This study may facilitate the quality of medicinal plants through gene engineering in the future.
