Cloning and functional analysis of IPI gene from Fritillaria unibracteata Hsiao et K. C. Hsia.
10.16438/j.0513-4870.2022-0925
- VernacularTitle:暗紫贝母IPI基因的克隆与功能分析
- Author:
Jiao CHEN
1
;
Si-min SONG
1
;
Jie TANG
1
;
Jin-xiu XIN
1
;
Qian ZHANG
1
;
Hong-jie ZHAO
1
;
Xin CHEN
2
;
Jia-yu ZHOU
1
;
Hai LIAO
1
Author Information
1. School of Life Science and Engineering, Southwest Jiaotong University, Chengdu 610031, China
2. The Third People's Hospital of Chengdu, Chengdu 610014, China
- Publication Type:Research Article
- Keywords:
italic>Fritillaria unibracteata;
isopentenyl-diphosphate delta isomerase;
gene cloning;
heterologous expression;
italic>in vitro activity
- From:
Acta Pharmaceutica Sinica
2023;58(2):447-453
- CountryChina
- Language:Chinese
-
Abstract:
An open reading frame (ORF) of isopentenyl-diphosphate delta isomerase gene (FuIPI) was cloned from Fritillaria unibracteata Hsiao et K. C. Hsia. (F. unibracteata). Furthermore, the bioinformatics and functional analyses of FuIPI were performed in this study. The result showed that, the ORF of FuIPI gene was 825 bp, encoding a polypeptide of 274 amino acids in length, with a relative molecular mass of about 31 kD and a theoretical isoelectric point of 5.61. Sequence analysis showed that FuIPI contained conserved structural domains and key residues involved in the catalyzing process. The phylogenetic analysis exhibited that FuIPI was closely related to IPIs of Dendrobium officinale and Musa acuminate. Real-time PCR analysis showed that FuIPI was distributed in different tissues of F. unibracteata, but had the highest transcriptional level in leaves, followed by stems, bulbs, and flowers. Furthermore, the FuIPI protein was successfully expressed in Escherichia coli BL21(DE3). The purified FuIPI protein successfully catalyzed the conversion from isopentenyl diphosphate (IPP) to dimethylallyl pyrophosphate (DMAPP). The above results provided a theoretical basis for further investigation of the molecular role of FuIPI in the biosynthesis of alkaloids.
