来自苏云金芽孢杆菌日本血清型菌株buibui的对金龟子昆虫有毒的一种新型晶体蛋白基因的克隆、异源表达及定位
Cloning, heterologous expression, and localization of a novel crystal protein gene from Bacillus thuringiensis serovar japonensis strain buibui toxic to scarabaeid insects.
作者信息
Sato R, Takeuchi K, Ogiwara K, Minami M, Kaji Y, Suzuki N, Hori H, Asano S, Ohba M, Iwahana H
机构信息
Department of Applied Biological Science, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Japan.
出版信息
Curr Microbiol. 1994 Jan;28(1):15-9. doi: 10.1007/BF01575980.
Recombinant Escherichia coli strains harboring pAG1, pAG2, pKBB100, and pKBB101 were cloned by using antiserum constructed against 130-kDa crystal protein antigen of Bacillus thuringiensis serovar japonensis strain Buibui. DNAs in the recombinant strains hybridized to the 26-base oligonucleotide probe corresponding to N-terminal amino acids of the 130-kDa crystal protein of strain Buibui. Cultures of the recombinant strains were toxic to larvae of the cupreous chafer, Anomala cuprea. Furthermore, the production of the 130-kDa polypeptide was demonstrated in the cells harboring pAG1 and pAG2 by immunoblot analysis with antiserum against the 130-kDa crystal protein. Southern hybridization analysis showed that the 130-kDa crystal protein gene is located on the chromosomal DNA of strain Buibui. On the other hand, DNA probes derived from cryIA(a) and cryIIIA genes did not hybridize to the DNA of strain Buibui.
利用针对苏云金芽孢杆菌日本变种Buibui菌株130 kDa晶体蛋白抗原制备的抗血清，克隆了携带pAG1、pAG2、pKBB100和pKBB101的重组大肠杆菌菌株。重组菌株中的DNA与对应于Buibui菌株130 kDa晶体蛋白N端氨基酸的26碱基寡核苷酸探针杂交。重组菌株的培养物对铜绿丽金龟幼虫有毒。此外，通过用针对130 kDa晶体蛋白的抗血清进行免疫印迹分析，在携带pAG1和pAG2的细胞中证实了130 kDa多肽的产生。Southern杂交分析表明，130 kDa晶体蛋白基因位于Buibui菌株的染色体DNA上。另一方面，源自cryIA(a)和cryIIIA基因的DNA探针未与Buibui菌株的DNA杂交。
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