[Separation of Bacteroides forsythus ATCC43037 proteins by horizontal two-dimensional gel electrophoresis].
OBJECTIVE
To set up a rapid, efficient, reliable and accurate method for separation of Bacteriodes forsythus proteins.


METHODS
Bacteroides forsythus ATCC43037 cells were harvested by centrifugation, washed in Tris buffer to remove excess medium, and lysed by sonication. The sonicated lysis proteins were extracted step by step with ReadyPrep Sequential Extraction Kit (Bio-Rad). The supernatant of B. forsythus proteins were used for two-dimensional gel electrophoresis. The first dimension IEF (isoelectric focusing) was run with Immobiline DryStrip (pH 3-10) and the second dimension SDS-PAGE was run with Excelgel SDS, gradient 8-18 precast gel and buffer strips. The separated proteins were stained with Coomassie Brilliant Blue and silver staining kit (Plusone Silver Staining Kit, Protein, Pharmacia Biotech).


RESULTS
1. The protein spots were clear when sample cups were used in the middle of IPG strip during IEF. 2. B. forsythus proteins were separated clearly by horizontal two-dimensional electrophoresis and most of B. forsythus whole-cell proteins were acidic proteins (P13-7).


CONCLUSIONS
Horizontal two-dimension electrophoresis is a useful method for separating B. forsythus proteins.
Key. Laboratory of Oral Biomedical Engineering Ministry of Education, Sichuan University, Chengdu 610041, China.
Ding-ming
DM
Huang
Xue-dong
XD
Zhou
Masae
M
Kuboniwa
Atso
A
Amano
D001426:Bacterial Proteins / Q000302:isolation & purification*; D001439:Bacteroides / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods
D001426:Bacterial Proteins