Cloning, induction pattern and prokaryotic expression of a small heat shock protein SmHSP21.8 gene from Salvia miltiorrhiza
10.16438/j.0513-4870.2021-1797
- VernacularTitle:丹参小分子热激蛋白SmHSP21.8基因克隆、诱导模式和原核表达
- Author:
Shi-wei WANG
1
;
Ren-jun QU
2
;
Jia-ming PENG
2
;
Xin-xin WANG
2
;
Chen-jing SHI
2
;
Han ZHENG
2
;
Ye SHEN
2
;
Lu-qi HUANG
2
Author Information
1. School of Traditional Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou 510006, China; State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
2. State Key Laboratory of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
- Publication Type:Research Article
- Keywords:
italic>Salvia miltiorrhiza;
small heat shock protein;
gene cloning;
prokaryotic expression;
stress treatment
- From:
Acta Pharmaceutica Sinica
2022;57(6):1909-1917
- CountryChina
- Language:Chinese
-
Abstract:
In order to reveal the molecular mechanism of the small heat shock proteins (sHSPs) involved in stress resistance and active ingredients accumulation in Salvia miltiorrhiza, a small heat shock protein gene was cloned from Salvia miltiorrhiza by reverse transcription PCR according to the transcriptome data of orange root Salvia miltiorrhiza. The gene is named SmHSP21.8 based on the molecular weight of the protein, and it contains an open reading frame of 585 bp, which encodes 194 amino acids. The results of phylogenetic analysis and amino acid sequence alignment showed that SmHSP21.8 protein belongs to the endoplasmic reticulum (ER) subfamily, and contains a conserved endoplasmic reticulum-specific DPFR-I/V-LE-H/Q-x-P motif at N-terminus. The prokaryotic expression vector pMAL-c2X-SmHSP21.8 was constructed and transformed into E. coli BL21 competent cells. The recombinant protein was successfully expressed after inducted. Temporal and spatial expression analysis showed that SmHSP21.8 gene was the highest expressed in flowers and had significant tissue specificity. The relative expression of the gene was significantly increased in seedlings after induction by 38 ℃, PEG6000, abscisic acid(ABA), and indole-3-acetic acid (IAA), indicating that SmHSP21.8 gene may be involved in abiotic stress such as high temperature and drought, as well as the response to exogenous hormones ABA and IAA. These results lay the foundation for further research on the molecular mechanism of small heat shock proteins involved in adversity stress.
