Preparation The Nanowires Of Sup35-Vip3A Fusion Protein And Genome Analysis Of Bacillus Thuringiensis Strain CT-43

1 Preparation the nanowires of Sup35-Vip3A fusion proteinThe vegetative insecticidal protein (Vip) is a kind of toxin proteins expressed in the vegetative stage of growth from the starting of mid-log phase to the sporulation phase in Bacillus thuringiensis cells, which is unlike the insecticidal crystal protein in B. thuringiensis, whose expression relies on sporulation. Vips were distributed widely in wild isolates of B. thuringiensis. Vip3Aa, one of Vip3A, was confirmed has especially high toxicity against Agrotis ipsilon and Spodoptera exigua.Sup35 is a translation termination factor in its native state in Saccharomyces cerevisiae. The prion domain of Sup35p can form amyloid-like proteinaceous fibrils both in vitro and in vivo. Because of its ability to self-assemble into nanowires, the prion domain of Sup35p has been widely used in biotechnology and nanotechnology.In this research, Vip3Aa and Vip3Aa with small fluorescent tag biarsenical-tetracysteine (TC) were linked to the yeast prion protein Sup351-61 respectively through the way of gene fusion. The results suggested that the fusion proteins Sup351-61-Vip3Aa and Sup351-61-Vip3Aa-TC could self-assemble into nanowires. Whether the fusion proteins can improve the insecticidal activities after the self-assembly to nanowire and how Vip3Aa is digested in insects are still on studying.2 Genome analysis of Bacillus thuringiensis strain CT-43B. thuringiensis subsp. chinensis strain CT-43 has high toxin against Lepidopterous and Dipterous insects. It can form various parasporal crystals consisting of various insecticidal crystal proteins containing CrylAa3, Cry1Bal, Cry1Ia14, Cry2Aa9, and Cry2Abl. During fermentation, it simultaneously generates vegetative insecticidal protein Vip3Aa10, as well as insecticidal nucleotide analogue thuringiensin.In this research, we classified the different kinds of genes refer to tanscription factor, sigma factor, phage, transposon, insertion sequence and spore germination according to genome annotation results of B. thuringiensis strain CT-43. At the same time, we find out 258 putative small RNA (sRNA) from the gene spacers by the use of bioinformatics and also preliminary analyze the secondary structures and nearby operons of 25 sRNA among them. These work could lay a foundation for our further study of the metabolism and control mechanisms in B. thuringiensis strain CT-43.