Establishment of extraction method and 2-dimensional electrophoresis conditions for root tuber proteome analysis of Rehmannia glutinosa.
- Author:
Linkun WU
1
;
Haibin WANG
;
Chuihuai YOU
;
Zhixing ZHANG
;
Miaomiao NIU
;
Zhongyi ZHANG
;
Wenxiong LIN
Author Information
- Publication Type:Journal Article
- MeSH: Electrophoresis, Gel, Two-Dimensional; methods; Plant Proteins; analysis; chemistry; Plant Roots; chemistry; Proteome; chemistry; isolation & purification; Rehmannia; chemistry
- From: China Journal of Chinese Materia Medica 2011;36(8):984-987
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish an efficient and high resolution 2-DE (two-dimensional electrophoresis) protocol for root tuber proteome analysis of Rehmannia glutinosa.
METHODProteins from root tuber of R. glutinosa were extracted by using five different methods and their productivity and profiles were assessed by means of SDS-PAGE and two-dimensional electrophoresis.
RESULTThe trichloroacetic acid (TCA)-phenol extraction method was found most effective for the extraction with the highest protein yield, the most spots in protein patterns, and the highest resolution of proteins, and the clearest background could be achieved simultaneously. A 1:5 solution of ampholine pH 3-10 and pH 5-8 for a nonlinear gel and the 170 microg of protein loading dosage obtained maps with more protein spots and higher resolution of separation patterns.
CONCLUSIONThis study based on the optimized root tuber proteome preparation and the 2-DE protocol gets a high resolution and reproducibility 2-DE image, which will be expected to have excellent applications in proteomics studies of R. glutinosa tuber root.
