Abstract:

Objective  To optimize the conditions of  two-dimensional electrophoresis (2-DE) and its related techniques in  research on proteomics of Proteus mirabilis, and to elucidate the effects of the supersonic power,the agitate time and the loading amount of proteins on 2-DE maps.Methods One strain of  Proteus mirabilis
 was chosen,and  its proteins were extracted by sonicating on a sonicator fitted with a microprobe.and the supersonic powers were 130 W and 80 W.The same condition of immobilized pH gradient isoelectric focusing and SDS-PAGE was used to separate the proteins obtained  from the two ways.1 and 2 mg of samples of the total  proteins obtained  from Proteus mirabilis by 80 W supersonic power were collected for  2-DE on 17 cm IPG strips under the same condition.Results  The 2-DE map of the proteins   obtained by 130 W supersonic power  was clear,repeatable and had more protein spots than that obtained by 80 W supersonic power.The 2-DE map with sample amount of 1 mg exhibited less vertical and horizontal stripes than that of 2 mg, which appeared good repeatability.Conclusion  Adopting lower supersonic power and a few of  samples can get better 2-DE map.
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Key words: Proteus mirabilis, proteomics；two-dimensional electrophoresis