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苏芸金芽孢杆菌的肠毒素及其编码基因的研究

张文成　任改新

　　【摘要】　目的　对苏芸金芽孢杆菌(简称Bt )种群不同亚种存在的蜡状芽孢杆菌肠毒素基因及溶血素BL进行检测，以便评估Bt对人类及高等动物潜在的致病性。方法　采用多重引物PCR技术，在同一PCR体系同时加两对引物，hblA及bceT基因的扩增片段分别为1017bp及427bp，与预期的完全一致；应用HBL羊血平板，产溶血素BL的菌株可产生非连续型溶血环。结果　45株Bt标准株中，29株含有hblA基因，占64.4%;15个生产用菌株及野生株中，含hblA基因的菌株为10株，占66.66%;含bceT基因的标准株29株，占64.4％，含bceT的Bt生产用菌株及野生株为13株，占86.6%。HBL羊血平板检测结果表明：28.8%的Bt标准株及33.3%的Bt生产用菌株及野生株含溶血素BL。结论　hblA及bceT基因在苏芸金芽孢杆菌中是普遍存在的，建议在遴选Bt生产菌株时，选用不含肠毒素基因或含肠毒素基因而不表达或表达水平不高的菌株。
　　【主题词】　聚合酶链反应　芽孢杆菌，苏芸金　芽孢杆菌，蜡状　肠毒素类　基因

Study of the bceT and hblA genes and the hemolysin BL of Bacillus thuringiensis group　Zhang Wencheng，Ren Gaixin. Department of Biology Nankai University，Tianjin 300071
　　【Abstract】　Objective　The hemolysin BL and the genes encoding enterotoxins have been examined in order to study the potential pathogenicity of Bt to mammals and human beings. Methods　Multiple polymerase chains reaction was performed to detect the hblA and bceT genes at the same time,the amplified fragments were 1071bp and 427bp,respectively,as we expected.The HBL blood agar was used to detect the hemolysin-producing Bt strains. Results　29 Bt type strains(64.4%) and 10 Bt commercial strains(66.6%) have the hblA gene; 29 Bt type strains(64.4%) and 13 Bt commercial strains(86.6%) have the bceT gene. Identification of hemolysin-producing Bt strains was conducted by the discontinuous hemolytic patterns on the HBL blood nutrient agar plates. 13 Bt type strains(28.8%) and 5 commercial strains(33.3%) produced the hemolysin BL. Conclusions　The majority of Bt strains have hblA and bceT genes. In order to avoid the hazards to human beings, we suggest that the Bt strains which have no the two genes or the strains don t express the above genes can be used as the master strains for producing insecticides.
　　【Subject words】　Polymerase chain reaction　Bacillus thuringiensis　Enterotoxin gene　Bacillus cereus

　　由于苏芸金芽孢杆菌（Bacillus thuringiensis,简称Bt)在其芽孢形成期可产生多种杀虫晶体蛋白，目前已成为全球性应用最广泛的细菌杀虫剂

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