苏云金芽胞杆菌肠毒素基因的PCR检测  

DETECTION OF ENTEROTOXIN GENES IN BACILLUS THURINGIENSIS BY PCR

作　　者：袁志明[1] 蔡全信[1] Andrup L Eilenberg J 庞义[4] 

机构地区：[1]中国科学院武汉病毒研究所,武汉430071 [2]丹麦国立职业健康研究所 [3]丹麦皇家畜牧和农业大学 [4]中山大学生物防治国家重点实验室,广州510275

出　　处：《微生物学报》2001年第2期148-154,共7页Acta Microbiologica Sinica

摘　　要：采用多重引物PCR进行了 45株苏云金芽胞杆菌、2株蜡状芽胞杆菌和 2株球形芽胞杆菌溶血素BL ,肠毒素T和entS基因的检测 ,结果表明 95 6%苏云金芽胞杆菌含溶血素hblA基因 ,91 1 %含bceT基因 ,93 3%含entS基因。用两种商业化肠毒素检测试剂盒TECRA和RPLA进行所有菌株肠毒素的体外免疫测定 ,大部分苏云金芽胞杆菌和阳性蜡状芽胞杆菌都能产生不同水平的肠毒素活性 ,同hblA基因PCR检测结果基本相符。尽管DBT0 0 7和T2 4 0 0 1含有hblA基因 ,但用TECRA却检测不到肠毒素 ;Dmu39菌株不含肠毒素基因 ,但用TECRA却检测出高的肠毒素活性。苏云金芽胞杆菌BDT2 4 8和球性芽孢杆菌不含肠毒素基因和肠毒素。The presence of hemolysin HblA( hbl A),enterotoxin BceT( bce T)and enterotoxin S( ent S)genes from 45 strains of B.thuringiensis, 4 strains of B.cereus and B.sphaericus have been detected respectively by multiple PCR.The results showed that 95 6% B.thuringiensis strains contain the B component of hblA gene,91 1% and 93 3% of them contain bceT and ent S genes sequences respectively.The enterotoxin productions in all strains have also been analysis using two commercial immunoassay kits(TECRA and RPLA)and it has proved that most of B.thuringiensis stains and the positive B.cereus strain can produce entero toxins during their growth.However,the two hblA sequence positive stains,DBT007 was negative when tested both by RPLA and Tecra,T24 001 was negative when assayed by Tecra and positive by RPLA.One hblA sequence negative strain Dmu39 was negative when tested by RPLA but positive by Tecra.No enterotoxin and enterotoxin gene could be detected in B.thuringiensis DBT248 and the B.sphaericus strains.The results suggest that the potential risk of using B.thuringiensis, as biopesticide needs to be further evaluated.

关 键 词：苏云金芽胞杆菌 蜡状芽胞杆菌 球性芽胞杆菌 肠毒素 检测 

分 类 号：Q785[生物学—分子生物学]