Proteomics Study Of Insecticidal Crystal Proteins From Bacillus Thuringiensis Strain 4.0718 And Subsp. Kurstaki HD-1

A novel Bacillus thuringiensis strain Bt 4.0718 selected in our laboratory presents higher toxicity and broader spectrum to insects in comparison with HD-1, in the apply, it also exhibits such characteristics. In order to compare the insecticidal crystal proteins from Bacillus thuringiensis strain 4.0718 and subsp. kurstaki HD-1, investigate the relationship between insecticidal crystal proteins and toxicity, and acquire insights of the mechanism of toxic specificity and the action mode of these toxins.A series of methods, including two dimensional gel electrophoresis, silver staining, PDQuest 2-DE software analysis, pepetide mass fingerprinting, SWISS-PROT database searching and CPHmodles, Ds ViewerPro Trial, Swiss-PdbViewer software analysis were used. By comparing and optimizing five kinds of lysis solution, the volume of sample loading and the protocol for isoelectric focusing, a well-focused 2-DE map with high resolution and reproducibility is achieved, protein profiles of Bt 4.0718 and HD-1 are similar, protein spots are mainly distributed in pH 4-7 and Mr 10-130 kDa, After silver staining, the 2-DE image analysis by PDQuest 2-DE software detects average 122 + 9 spots in Bt 4.0718, and 126+11 spots in HD-1. There are 64 matched spots between Bt 4. 0718 and HD-1 maps, 8 spots are only detected in HD-1, and 6 spots are only detected in Bt 4.0718, 11 protein spots are found to increase or decrease between Bt 4.0718 and HD-1. it indicates the similarity and variability in toxicity of Bt 4.0718 and HD-1. A part of spots are cut off from silver staining gel, digested...