Preliminary Study On The Differential Electrophoresis Proteomics Analysis Of The Resistant And Susceptible Trichoplusia Ni Cells To Bt Cry1Ac Toxin

In 1957 American Pacific Corporation Yeast produced Bacillus thuringiensis commercial products, it had played an important role in agriculture, forestry and pest control as the microbial pesticides. It was safe to human beings and animals, pollution-free, high yield and good efficiency, and it had become the most successful biological pesticide in use at home and abroad. However, pest had already had a noticeable resistance to them, with its extensive and diversified use and the promotion of transgenic Bt plants It had became an important issue that how to knock-down their resistance to Bt. Only clarifying the molecular mechanisms could we draw effective resistance management strategy and delay the development of resistance.The early laboratory work had been proved that Trichoplusia ni cells in vitro were able to produce high level of resistance to Bt crystal protein Cry1Ac.We took BT1-TN-5B1 cells, which was sensitive to Bt Cry1Ac toxin, and homologous resistance cells as the materials.When the cells were treated with Cry1Ac continually and high-intensity, we revealed the differences betwteen them, using two-dimensional gel electrophoresis and mass spectrometry technology.By the two-dimensional electrophoresis technology, we got the proteomics two-dimensional electrophoresis maps in both cells, and analyzed with ImageMaster Elite 5.0v software. About 896 dots in average were detected on the resistant cells' proteomics 2-DE maps, and about 838 dots on the susceptible cells'. Most of the protein spots distributed between 14KD and 100KD, and pI between pH4 and pH9.We excavated several protein spots with good repeatability from the gels.These spots were identified through the Peptide mass fingerprint spectrumtry and Mascot, and several proteins were matched in current protein database.From the PMF maps, the closest protein in scores of peptides and the coverage rate was a flagellar motor protein in proteobacteriaγ-proteobacteria Idiomarina loihiensis L2TR, it had a certain degree of homology with the bacterial membrane protein. It showed that the unknow protein might be a kind of membrane protein associated with the flagellar motor protein.In addition, the peptide analysis of the spot on PMF map showed that 1st to 44th peptides were perfectly matched with Formamidopyrimidine-DNA glycosylase, next we could identified the protein with N-terminal sequence testing.But the other one was failed to get the right match with any protein. And a differential spot on the susceptible cells'gel was also analyzed, the results showed that it was homologous to lipopolysaccharide biosynthesis protein.They might be related to the development of Bt resistance.