Dissolution of Expolysaccharide and Two-Dimensional Electrophoresis Construction of Bacillus mucilaginosus

Abstract:

In order to establish two-dimensional electrophoresis（2-DE） technology system of total proteins of Bacillus mucilaginosus，2-DE maps obtained from different microbial expolysaccharide dissolution methods，protein extraction methods，pH ranges of IPG strips and isoelectric focusing（IEF） programs were compared. The results showed that hydrochloric acid solution could dissolute expolysaccharide effectively，and phenol extraction protocol combined with TCA/acetone was more beneficial to total protein extraction from Bacillus mucilaginosus. The 2-DE maps without obvious vertical and horizontal stripes，with low background，high resolution and good reproducibility were obtained using the following optimized parameters：17 cm IPG strip with pH 4~7，program Ⅱ（100 V 30 min，1000 V 3 h，8000 V 5 h，8000 V 60000 Vh，500 V 20 h）. Analyzed by the PDQuest software，about 700 protein spots were isolated from the 2-DE protein map. The construction of 2-DE technology system will make a major contribution to the further study of B. mucilaginosus.