Cloning and protein expression analysis of geranyl diphosphate synthase genes in Tripterygium wilfordii.
- Author:
Li-Chan TU
1
;
Yi-Feng ZHANG
1
;
Ping SU
1
;
Tian-Yuan HU
1
;
Yu-Ru TONG
1
;
Hong-Yu GUAN
1
;
Yu-Jun ZHAO
2
;
Xia-Nan ZHANG
1
;
Yuan YUAN
2
;
Wei GAO
1
;
Lu-Qi HUANG
2
Author Information
- Publication Type:Journal Article
- Keywords: Tripterygium wilfordii; bioinformatics analysis; cloning; geranyl diphosphate synthase; protein expression
- From: China Journal of Chinese Materia Medica 2017;42(2):220-225
- CountryChina
- Language:Chinese
- Abstract: Based on the transcriptome data, the study cloned full-length cDNA of TwGPPS1 and TwGPPS2 genes from Tripterygium wilfordii suspension cells and then analyzed the bioinformation of the sequence and protein expression. The cloned TwGPPS1 has a 1 278 bp open reading frame (ORF) encoding a polypeptide of 425 amino acids. The deduced isoelectric point (pI) was 6.68, a calculated molecular weight was about 47.189 kDa. The full-length cDNA of the TwGPPS2 contains a 1 269 bp open reading frame (ORF) encoding a polypeptide of 422 amino acids. The deduced isoelectric point (pI) was 6.71, a calculated molecular weight was about 46.774 kDa.The entire reading frame of TwGPPS1,2 was cloned into the pET-32a(+) vector and expressed in E. coli BL21 (DE3) cells to obtain the TwGPPS protein, which laid a basis for further study on the regulation of terpenoid secondary metabolism and biological synthesis.
