Ogiwara K, Indrasith L S, Asano S, Hori H
Laboratory of Biochemistry and Genetic Engineering, Tsukuba Laboratories of Research and Development, Ibaraki, Japan.
J Invertebr Pathol. 1992 Sep;60(2):121-6. doi: 10.1016/0022-2011(92)90084-h.
Midgut juices were prepared from Adoxophyes sp., smaller tea tortrix (STT); Bombyx mori, silkworm (SW); Spodoptera litura, common cutworm (CCW); Plutella xylostella, diamondback moth (DBM); and Musca domestica, housefly (HF) and immobilized onto Sepharose 4B. delta-Endotoxins (ICPs) from Bacillus thuringiensis subsp. kurstaki HD-1 and HD-73 were digested by these immobilized gut juice proteases. All gut juices tested derived relatively proteolytic resistant cores from ICP. The molecular sizes of these cores, about 55 kDa in SDS-PAGE, were resulted. In the case of CCW, however, digestion was very strong and only 1/20 concentration of core protein remained relative to other digests. The N-terminal amino acid sequencing of the core proteins showed that they were truncated at the very end of the N-terminus of protoxin, CryIA, at different sites. Although housefly larvae were completely insensitive to active toxin, the gut juice produced the core, suggesting that the housefly may lack the binding sites for the core-active toxin.
中肠汁液取自茶小卷叶蛾(Adoxophyes sp.,简称小茶卷叶蛾,STT)、家蚕(Bombyx mori,SW)、斜纹夜蛾(Spodoptera litura,CCW)、小菜蛾(Plutella xylostella,DBM)和家蝇(Musca domestica,HF),并固定在琼脂糖4B上。苏云金芽孢杆菌库尔斯塔克亚种HD-1和HD-73的δ-内毒素(ICP)被这些固定化的肠液蛋白酶消化。所有测试的肠液都能从ICP中产生相对抗蛋白水解的核心片段。在SDS-PAGE中,这些核心片段的分子大小约为55 kDa。然而,对于斜纹夜蛾来说,消化作用非常强烈,相对于其他消化产物,核心蛋白的浓度仅为1/20。核心蛋白的N端氨基酸测序表明,它们在原毒素CryIA的N端末端不同位点被截断。尽管家蝇幼虫对活性毒素完全不敏感,但肠液仍能产生核心片段,这表明家蝇可能缺乏核心活性毒素结合位点。