Establishment and optimization of two-dimensional gel electrophoresis of total proteins from Bacillus lincheniformis

A two dimensional gel electrophoresis protocol proteomic study of Bacillus lincheniformis and offer further information how Bacillus cereus stimulate the growth of Gluconobacter oxydans to produce 2-keto-L- gulonic acid ( 2-KLG) after entering into stationary phase was established.Different parameters, including protein preparation by different ultrasonic broken time, new type lysis, pH gradient and different sample of Bacillus lincheniformis protein, were used to evaluate the effect of two - dimensional electrophoresis of Bacillus lincheniformis proteins. Results showed that the clear background and reproducibility of two dimensional gel electrophoresis were established on the condition of using 15min of ultrasonic broken extraction, selection of protein cleavage I, pH4 ～ 7 24cm IPG strips, 80μg loading volume, isoelectric focus at 60V 15min, 120V 6h.The aim of this study was not only to explore a novel cell lysate in two- dimensional gel electrophoresis, but also to establish a method of proteome analysis for two-dimensional electrophoresis of Bacillus licheniformis.