Construction of yeast two-hybrid library of Salvia miltiorrhiza and screening of SmJAZ8 interaction protein.
10.19540/j.cnki.cjcmm.20180105.018
- Author:
Meng-Dan YANG
1
;
Ying ZHAO
2
;
Peng-da MA
1
Author Information
1. College of Life Science， Northwest A & F University， Yangling 712100， China.
2. College of Science， Northwest A & F University， Yangling 712100， China.
- Publication Type:Journal Article
- Keywords:
Salvia miltiorrhiza;
homogenization;
sequence analysis;
yeast two-hybrid
- MeSH:
Co-Repressor Proteins;
genetics;
DNA, Complementary;
Gene Library;
Plant Proteins;
genetics;
Salvia miltiorrhiza;
genetics;
Two-Hybrid System Techniques
- From:
China Journal of Chinese Materia Medica
2018;43(3):484-492
- CountryChina
- Language:Chinese
-
Abstract:
The study is aimed to construct high quality Salvia miltiorrhiza cDNA library and obtain the SmJAZ8 gene of S. miltiorrhiza by yeast two-hybrid system. In this study， full-length cDNA was synthesized from roots， stems， leaves， flowers and hairy roots of S. miltiorrhiza. The full-length cDNA library was synthesized by SMART method and constructed with DSN homogenization technique. The results showed that the library capacity was 1.45×10⁶， the recombination rate was 100%， and the average size of the insert was 500-2 000 bp. The recombinant vector of pDEST-pGADT7-SmJAZ8 was constructed and transformed into Y2HGold strain. The interaction protein was screened by yeast two-hybrid system. The DnaJ protein and UBQ protein were screened by yeast two-hybrid system. This study has successfully constructed a full-length cDNA library of S. miltiorrhiza， and laid the foundation for the follow-up study on functional gene screening and gene function of S. miltiorrhiza.
