Cloning, subcellular localization, and heterologous expression of ApNAC1 gene from Andrographis paniculata.
10.19540/j.cnki.cjcmm.20170217.005
- Author:
Jian WANG
1
;
Meng-Die QI
2
;
Juan GUO
3
;
Ye SHEN
3
;
Hui-Xin LIN
3
;
Lu-Qi HUANG
3
Author Information
1. College of Pharmacy, Anhui University of Chinese Medicine, Hefei 230038, China.
2. School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
3. State Key Laboratory of Dao-di Herbs Breeding Base, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
- Publication Type:Journal Article
- Keywords:
Andrographis paniculata;
NAC transcription factor;
expression pattern;
heterologous expression;
subcellular localization
- From:
China Journal of Chinese Materia Medica
2017;42(5):890-895
- CountryChina
- Language:Chinese
-
Abstract:
Andrographis paniculata is widely used as medicinal herb in China for a long time and andrographolide is its main medicinal constituent. To investigate the underlying andrographolide biosynthesis mechanisms, RNA-seq for A. paniculata leaves with MeJA treatment was performed. In A. paniculata transcriptomic data, the expression pattern of one member of NAC transcription factor family (ApNAC1) matched with andrographolide accumulation. The coding sequence of ApNAC1 was cloned by RT-PCR, and GenBank accession number was KY196416. The analysis of bioinformatics showed that the gene encodes a peptide of 323 amino acids, with a predicted relative molecular weight of 35.9 kDa and isoelectric point of 6.14. To confirm the subcellular localization, ApNAC1-GFP was transiently expressed in A. paniculata protoplast. The results indicated that ApNAC1 is a nucleus-localized protein. The analysis of real-time quantitative PCR revealed that ApNAC1 gene predominantly expresses in leaves. Compared with control sample, its expression abundance sharply increased with methyl jasmonate treatment. Based on its expression pattern, ApNAC1 gene might involve in andrographolide biosynthesis. ApNAC1 was heterologously expressed in Escherichia coli and recombinant protein was purified by Ni-NTA agarose. Further study will help us to understand the function of ApNAC1 in andrographolide biosynthesis.
