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通过Southern杂交发现高毒力苏云金芽胞杆菌 (Bacillusthuringiensis)YBT 1 52 0菌株含有两个杀虫晶体蛋白基因片段 ,其 5’ 末端所在HindⅢ片段分别为 6 8kb和 4 6kb ,它们对应的基因分别命名为cry2 1 8和cry4 6。经PCR鉴定 ,该菌含有cry1Aa、cry1Ab和cry1Ac基因 ,以及cry2基因 ,其中cry2 1 8属于cry1Ac。分析了cry2 1 8基因 41 90bp的核苷酸序列 ,在杀虫晶体蛋白基因分类系统中被命名为cry1Ac1 0。结合Southern杂交和PCR结果可判断 3个cry1A基因的拷贝数不同 ,其中cry1Ac拷贝数最高 ,YBT 1 52 0菌株与其它库斯塔克亚种的杀虫晶体蛋白基因所在限制性内切酶位置明显不同。
High-virulent Bacillus thuringiensis YBT1 52 0 strain was found to contain two insecticidal crystal protein gene fragments by Southern hybridization. The HindIII fragments at the 5 ’end of the strain were respectively 68 kb and 46 kb, and their corresponding genes were named as cry2 1 8 and cry4 6. The PCR identification, the bacteria contain cry1Aa, cry1Ab and cry1Ac gene, and cry2 gene, cry218 which belongs to cry1Ac. The 41 90 bp nucleotide sequence of cry2 1 8 gene was analyzed and named cry1Ac1 0 in the insecticidal crystal protein gene classification system. The results of Southern blotting and PCR showed that the copy number of three cry1A genes was different, among which the cry1Ac copy number was the highest, and the position of the restriction endonuclease of the insecticidal crystal protein gene of YBT1 52 0 strain and other Kustaki subspecies was obviously different.