Wasano N, Kim K H, Ohba M
Institute of Biological Control, Kyushu University, Fukuoka, Japan.
J Appl Microbiol. 1998 Apr;84(4):501-8. doi: 10.1046/j.1365-2672.1998.00371.x.
Four Lepidoptera-specific reference strains of Bacillus thuringiensis, belonging to serovars sumiyoshiensis (H3a:3d), fukuokaensis (H3a:3d:3e), darmstadiensis (H10a:10b) and japonensis (H23), which produce spherical parasporal inclusions, were examined for comparative characterization of delta-endotoxins. SDS-PAGE profiles of the alkali-solubilized parasporal inclusions revealed the presence of single major protein bands of 130 kDa in the four strains. Chymotrypsin and trypsin treatment of the proteins gave profiles different from those of the strains HD-1 (serovar kurstaki, H3a:3b:3c) and T84 A1 (serovar sotto, H4a:4b). Also, minor variations were observed in proteolysis profiles among the four strains. The LC50 values of purified parasporal inclusions for the silkworm (Bombyx mori) larvae were 7.35, 6.45, 3.08 and 2.63 micrograms g-1 diet, respectively, showing that their toxicity levels were 5-15 times lower than that of the strain HD-1 (0.49 microgram g-1 diet). Analysis by immunodiffusion and immunoblotting with polyclonal antisera revealed that parasporal inclusion proteins of the four strains are highly related, whereas they shared few or no common antigens with those of the strains HD-1, T84 A1 and Buibui (serovar japonensis).
对4种苏云金芽孢杆菌的鳞翅目特异性参考菌株进行了δ-内毒素的比较特性研究，这4种菌株分别属于sumiyoshiensis血清型（H3a:3d）、fukuokaensis血清型（H3a:3d:3e）、darmstadiensis血清型（H10a:10b）和japonensis血清型（H23），它们产生球形伴孢晶体。碱溶性伴孢晶体的SDS-PAGE图谱显示，这4种菌株均存在一条130 kDa的主要蛋白条带。用胰凝乳蛋白酶和胰蛋白酶处理这些蛋白质后得到的图谱与HD-1菌株（kurstaki血清型，H3a:3b:3c）和T84 A1菌株（sotto血清型，H4a:4b）不同。此外，在这4种菌株的蛋白水解图谱中也观察到了细微差异。纯化后的伴孢晶体对家蚕（Bombyx mori）幼虫的LC50值分别为7.35、6.45、3.08和2.63微克/克饲料，表明它们的毒性水平比HD-1菌株（0.49微克/克饲料）低5-15倍。用多克隆抗血清进行免疫扩散和免疫印迹分析表明，这4种菌株的伴孢晶体蛋白高度相关，而它们与HD-1、T84 A1和Buibui菌株（japonensis血清型）的伴孢晶体蛋白几乎没有共同抗原。
