Cloning and analysis of STR gene and its promoter from Uncaria
10.16438/j.0513-4870.2021-1700
- VernacularTitle:钩藤STR基因及其启动子的克隆与分析
- Author:
Hao ZHOU
;
Xing-xing LU
;
Wen-wen AO
;
Hai-min LIAO
;
Ming-sheng ZHANG
;
Wei QIANG
- Publication Type:Research Article
- Keywords:
italic>Uncaria rhynchophylla;
strictosidine synthase;
promoter;
subcellular localization;
prokaryotic expression
- From:
Acta Pharmaceutica Sinica
2022;57(5):1526-1536
- CountryChina
- Language:Chinese
-
Abstract:
On the basis of the Uncaria transcriptome, specific primers were designed for UrSTR. The full-length cDNA of UrSTR (GeneBank: OL310251) was 1 541 bp, encoding 345 amino acid residues, and the promoter region sequence of UrSTR (GeneBank: OL310252) was 1 179 bp. Phylogenetic tree is revealed that UrSTR had a closest relationship with STR from Ophiorrhiza pumila and Ophiorrhiza japonica. Localization of UrSTR protein is revealed located in the vacuole membrane. Plant-care analysis indicated that the promoter region sequence of UrSTR, covering multiple light, stress and hormone-response cis-regulatory elements, and verified transcriptional activity. The results of SDS-PAGE show that pET-28a-UrSTR recombinant protein was successfully expressed, and the size was anticipated. The UrSTR prokaryotic expression system needs to be optimized in the later stage. The research lays the foundation for further purification to study its structure and functional characterization of the UrSTR protein.
