Gill S S, Hornung J M, Ibarra J E, Singh G J, Federici B A
Appl Environ Microbiol. 1987 Jun;53(6):1251-6. doi: 10.1128/aem.53.6.1251-1256.1987.
The parasporal bodies of the mosquitocidal isolates of Bacillus thuringiensis subsp. israelensis and B. thuringiensis subsp. morrisoni isolate PG-14 were compared with regard to their hemolytic and cytolytic activities and the immunological relatedness of the 28- and 65-kilodalton (kDa) proteins that occur in both subspecies. The alkali-solubilized parasporal bodies of B. thuringiensis subsp. israelensis caused 50% lysis of human erythrocytes at 1.14 micrograms/ml, whereas those of B. thuringiensis subsp. morrisoni caused similar lysis at 1.84 micrograms/ml. Preincubation of solubilized parasporal bodies with dioleolyl phosphatidylcholine significantly inhibited the hemolytic activity of both supspecies. In cytolytic assays against Aedes albopictus cells, the toxin concentrations causing 50% lysis for B. thuringiensis subsp. israelensis and B. thuringiensis subsp. morrisoni were 1.87 and 11.98 micrograms/ml, respectively. Polyclonal antibodies raised separately against the 25-kDa protein (a tryptic digest of the 28-kDa protein) or the 65-kDa protein of B. thuringiensis subsp. israelensis cross-reacted, respectively, with the 28- and the 65-kDa proteins of B. thuringiensis subsp. morrisoni. However, neither of these antibodies cross-reacted with the 135-kDa protein of either subspecies. These results indicate that the mosquitocidal and hemolytic properties of B. thuringiensis subsp. israelensis and B. thuringiensis subsp. morrisoni isolate PG-14 are probably due to the biologically related proteins that are present in the parasporal bodies of both subspecies. The lower hemolytic activity of the B. thuringiensis subsp. morrisoni may be due to the presence of lower levels of the 28-kDa protein in that subspecies.(ABSTRACT TRUNCATED AT 250 WORDS)
对苏云金芽孢杆菌以色列亚种和苏云金芽孢杆菌莫里森氏亚种PG - 14的杀蚊分离株的伴胞晶体,就其溶血和细胞溶解活性以及两个亚种中均存在的28千道尔顿(kDa)和65千道尔顿蛋白质的免疫相关性进行了比较。苏云金芽孢杆菌以色列亚种经碱溶解的伴胞晶体在浓度为1.14微克/毫升时可使50%的人红细胞发生裂解,而苏云金芽孢杆菌莫里森氏亚种的伴胞晶体在浓度为1.84微克/毫升时产生类似程度的裂解。将溶解的伴胞晶体与二油酰磷脂酰胆碱预孵育可显著抑制两个亚种的溶血活性。在针对白纹伊蚊细胞的细胞溶解试验中,苏云金芽孢杆菌以色列亚种和苏云金芽孢杆菌莫里森氏亚种导致50%细胞裂解的毒素浓度分别为1.87微克/毫升和11.98微克/毫升。分别针对苏云金芽孢杆菌以色列亚种的25千道尔顿蛋白质(28千道尔顿蛋白质的胰蛋白酶消化产物)或65千道尔顿蛋白质产生的多克隆抗体,分别与苏云金芽孢杆菌莫里森氏亚种的28千道尔顿和65千道尔顿蛋白质发生交叉反应。然而,这些抗体均未与任一亚种的135千道尔顿蛋白质发生交叉反应。这些结果表明,苏云金芽孢杆菌以色列亚种和苏云金芽孢杆菌莫里森氏亚种PG - 14的杀蚊和溶血特性可能归因于两个亚种伴胞晶体中存在的生物学相关蛋白质。苏云金芽孢杆菌莫里森氏亚种较低的溶血活性可能是由于该亚种中28千道尔顿蛋白质的含量较低。(摘要截短于250字)