Quantitative Analysis Of Differential Proteins Between The Susceptible And Resistant Trichoplusia Ni Cells To Bt Cry1Ac Toxin In Culture

Since the 1950s, because of high toxicity, not killing predators and beneficial microorganisms, not destroying the ecological balance, to a large-scale fermentation, low-cost advantages, Bacillus thuringiensis pesticide has become the world's most widely used microbial pesticides. However, with the long-term and large-scale use ,most pests have had resistance to Bt,and the resistance showed a progressive trend. How to deal with Bt insect resistance in pest control has become an important issue, we must clarify its formation mechanism as soon as possible.From long term ,our laboratory has done a lot of research in this area, particularly against Trichoplusiani BTI-Tn-5B1-4 cells to produce high-level resistance to Cry1Ac toxin .And from the level of proteomics,we talk about the molecular mechanisms of resistance.Using two dimentional electrophoresis ,predecessors get some different protein spots by separating the total protein of the two sensitive and resistant cells and the membrane protein, silver staining color, Melanie Viewer II software analysis. We identify some distinct proteins by peptide mass fingerprint ( called PMF) and a protein data bank roughly.And maybe these proteins are homologous proteins relate to the peripheral cytoplasm protein, protein of lipopolysaccharide (LPS) biological synthesis and immunoglobulin heavy chain.This study is based on the work of toxicity mensuration,filtration and stability of cell resistance, studying differences in gene expression and identification of different proteins, using two-dimention electrophoresis to separate the total proteins of the sensitive and resistant cells , silver staining color, ImageMaster 2D Platinum 6.0 software analysis, statistics of the gel on all the differences in protein spots. Referring to 2D map of Trichoplusiani cells BTI-Tn-5B1-4 cell ,we found 209 different protein dots, among which the expression increase of two to three times is 17, the expression increase of more than three times is 18, here the expression increase of more than five times is four, the decrease is 13 and 7 homologously; Referring to 2D map of resistant cell,214 different spots are detected. The expression increase of two to three times is 13, the expression increase of more than three times is 10,the expression increase of more than five times is 3; the decrease is 17,28 and 12 homologously. With wonderful repeatability and good significance,we can get 11 dots by our eyes easily.Maybe these differences have a direct or indirect relationship to resistance formation.