Construction of Panax ginseng root yeast library and screening of interacting proteins of PgD14 and Pgpht2-1.
10.19540/j.cnki.cjcmm.20240813.102
- Author:
Hao LIANG
1
;
Hai SUN
1
;
Cai SHAO
1
;
Bo-Chen LYU
1
;
Wei-Yu CAO
1
;
Hong-Ju LONG
1
;
Ya-Yu ZHANG
2
Author Information
1. Jilin Provincial Key Laboratory of Traditional Chinese Medicinal Materials Cultivation and Propagation, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences Changchun 130112, China.
2. Jilin Provincial Key Laboratory of Traditional Chinese Medicinal Materials Cultivation and Propagation, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences Changchun 130112, China College of Food and Biological Engineering, Chengdu University Chengdu 610106, China.
- Publication Type:Journal Article
- Keywords:
Panax ginseng;
interacting protein;
yeast one-hybrid;
yeast two-hybrid
- MeSH:
Panax/metabolism*;
Two-Hybrid System Techniques;
Plant Proteins/metabolism*;
Gene Library;
Plant Roots/chemistry*;
Protein Binding;
Transcription Factors/metabolism*;
Promoter Regions, Genetic
- From:
China Journal of Chinese Materia Medica
2024;49(22):6107-6118
- CountryChina
- Language:Chinese
-
Abstract:
To construct a high-quality Panax ginseng cDNA library, transcription factors binding to the P. ginseng PgD14 gene promoter were screened by yeast one-hybrid, and proteins interacting with the P. ginseng Pgpht2-1 gene-encoded protein were screened by yeast two-hybrid. In this study, root tissues of P. ginseng were used as materials. Gateway technology was used to construct the P. ginseng yeast one-hybrid library, and duplex-specific nuclease(DSN) homogenization technology was used to construct the P. ginseng yeast two-hybrid library. The pAbAi-PgD14-Pro961 vector was used as bait to screen candidate transcription factors that might bind to the PgD14 gene promoter from the yeast one-hybrid library, and the pGBKT7-Pgpht2-1 vector was used as bait to screen candidate proteins that might interact with the Pgpht2-1 gene-encoded protein from the yeast two-hybrid library. The yeast one-hybrid library had a size of 1.20×10~7 CFU, a recombination rate of 100%, and an average inserted fragment length of more than 1 000 bp. The yeast two-hybrid library had a size of 1.832×10~5 CFU, a recombination rate of 100%, and an average inserted fragment length of about 1 000 bp. The recombinant vectors pAbAi-PgD14-Pro961 and pGBKT7-Pgpht2-1 were transformed into Y1HGold and AH109 strains, respectively, and interacting proteins were screened by yeast one-hybrid and yeast two-hybrid. As a result, 54 transcription factors that could bind to the PgD14 gene promoter of P. ginseng and 42 proteins that may interact with the protein encoded by the Pgpht2-1 gene were identified. This study successfully constructed the P. ginseng yeast one-hybrid and yeast two-hybrid cDNA libraries, laying a foundation for subsequent studies on the functions of the P. ginseng PgD14, Pgpht2-1, and other genes.
