Monoclonal Antibody Analysis and Insecticidal Spectrum of Three Types of Lepidopteran-Specific Insecticidal Crystal Proteins of Bacillus thuringiensis
Monoclonal Antibody Analysis and Insecticidal Spectrum of Three Types of Lepidopteran-Specific Insecticidal Crystal Proteins of Bacillus thuringiensis
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苏云金芽孢杆菌三种鳞翅目特异性杀虫晶体蛋白的单克隆抗体分析及杀虫谱
DOI：
10.1128/aem.54.8.2010-2017.1988
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发表时间：
1988
影响因子：
4.4
通讯作者：
M. Vaeck
中科院分区：
文献类型：
--
作者：
H. Höfte;J. Van Rie;S. Jansens;A. Van Houtven;H. Vanderbruggen;M. Vaeck
我们研究了 29 种对鳞翅目幼虫具有活性的苏云金芽孢杆菌晶体的蛋白质组成和杀虫谱。所有晶体均含有 130 至 140 千道尔顿 (kDa) 的蛋白质，根据原毒素和胰蛋白酶激活的核心片段的分子量可将其分为三种类型。当针对五种鳞翅目物种进行测试时，这三种类型的蛋白质显示出特有的杀虫谱。 A型晶体蛋白是130或133 kDa的原毒素，经胰蛋白酶加工成60 kDa的毒素。编码这种类型晶体蛋白的几个基因较早已被克隆和测序。它们在有毒片段的 N 端一半高度保守，之前根据其基因的限制性图谱将其分为三个亚型（4.5、5.3 和 6.6 千碱基亚型）。目前的研究表明，这三种亚型的不同蛋白质对天蛾和菜粉蝶具有相同的毒性，并且对灰翅夜蛾没有可检测到的活性。然而，4.5、5.3 和 6.6 千碱基亚型对烟夜蛾和甘蓝夜蛾的毒性不同。 B 型晶体蛋白由 140 kDa 的原毒素和 55 kDa 的胰蛋白酶核心片段组成。这些药物仅对所测试的五种昆虫之一（十字花科昆虫）有活性。 C 型原毒素和胰蛋白酶激活毒素分别为 135 kDa 和 63 kDa 蛋白质。这种类型的蛋白质与对北沙门氏菌和芸苔小球藻的高毒性有关。使用一组 35 种单克隆抗体来比较三种不同类型和亚型的晶体蛋白的结构特征。每种类型的蛋白质都可能与典型的表位结构相关，表明抗原结构和昆虫特异性之间存在明确的相关性。
We have investigated the protein composition and the insecticidal spectrum of crystals of 29 Bacillus thuringiensis strains active against lepidopteran larvae. All crystals contained proteins of 130 to 140 kilodaltons (kDa) which could be grouped into three types by the molecular weight of the protoxin and the trypsin-activated core fragment. Proteins of the three types showed a characteristic insecticidal spectrum when tested against five lepidopteran species. Type A crystal proteins were protoxins of 130 or 133 kDa, which were processed into 60-kDa toxins by trypsin. Several genes encoding crystal proteins of this type have been cloned and sequenced earlier. They are highly conserved in the N-terminal half of the toxic fragment and were previously classified in three subtypes (the 4.5-, 5.3-, and 6.6-kilobase subtypes) based on the restriction map of their genes. The present study shows that different proteins of these three subtypes were equally toxic against Manduca sexta and Pieris brassicae and had no detectable activity against Spodoptera littoralis. However, the 4.5-, 5.3-, and 6.6-kilobase subtypes differed in their toxicity against Heliothis virescens and Mamestra brassicae. Type B crystal proteins consisted of 140-kDa protoxins with a 55-kDa tryptic core fragment. These were only active against one of the five insect species tested (P. brassicae). The protoxin and the trypsin-activated toxin of type C were 135- and 63-kDa proteins, respectively. Proteins of this type were associated with high toxicity against S. littoralis and M. brassicae. A panel of 35 monoclonal antibodies was used to compare the structural characteristics of crystal proteins of the three different types and subtypes. Each type of protein could be associated with a typical epitope structure, indicating an unambiguous correlation between antigenic structure and insect specificity.