Construction of yeast one-hybrid library and screening of transcription factors regulating LS expression in Ganoderma lucidum.
10.19540/j.cnki.cjcmm.20190730.102
- Author:
Xiao-Lan XU
1
;
Feng-Li ZHU
2
;
Rong-Cai LAI
1
;
Lin-Chun SHI
3
;
Shi-Lin CHEN
4
Author Information
1. College of Bee Science,Fujian Agriculture and Forestry University Fuzhou 350002,China.
2. College of Food science,Fujian Agriculture and Forestry University Fuzhou 350002,China.
3. Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences,Peking Union Medical College Beijing 100193,China.
4. Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences Beijing 100700,China.
- Publication Type:Journal Article
- Keywords:
Ganoderma lucidum;
ganoderma triterpenoid;
regulatory factors;
yeast one-hybrid
- MeSH:
Gene Library;
Intramolecular Transferases/metabolism*;
Reishi/genetics*;
Saccharomyces cerevisiae;
Transcription Factors/metabolism*
- From:
China Journal of Chinese Materia Medica
2019;44(18):3967-3973
- CountryChina
- Language:Chinese
-
Abstract:
Lanosterol synthase( LS) is a key enzyme involving in the mevalonate pathway( MVA pathway) to produce lanosterol,which is a precursor of ganoderma triterpenoid. And the transcriptional regulation of LS gene directly affects the content of triterpenes in Ganoderma lucidum. In order to study the transcriptional regulation mechanism of LS gene,yeast one-hybrid technique was used to screen the transcription regulators which interact withthe promoter of LS. The bait vector was constructed by LS promoter,then the vector was transformed yeast cells to construct bait yeast strain. One-hybrid c DNA library was constructed via SMART technology. Then the c DNA and p GADT7-Rec vector were co-transformed into the bait yeast strain to screen the upstream regulatory factors of the promoter region of LS by homologous recombination. Total of 23 positive clones were screened. After sequencing,blast was performed against the whole-genome sequence of G. lucidum. As a result,8 regulatory factors were screened out including the transcription initiation TFIIB,the alpha/beta hydrolase super family,ALDH-SF superfamily,60 S ribosomal protein L21,ATP synthase β-subunit,microtubule associated protein Cript,prote asome subunit β-1,and transaldolase. Until now,the regulation effect of these 8 regulatory factors in G.lucidum has not been reported. This study provides candidate proteins for in-depth study on the expression regulation of LS.
