Servant P, Rosso M L, Hamon S, Poncet S, Del cluse A, Rapoport G
Unité de Biochimie Microbienne, URA 1300 du Centre National de la Recherche Scientifique, Institut Pasteur, 75724 Paris, France.
Appl Environ Microbiol. 1999 Jul;65(7):3021-6. doi: 10.1128/AEM.65.7.3021-3026.1999.
Cry11A from Bacillus thuringiensis subsp. israelensis and Cry11Ba from Bacillus thuringiensis subsp. jegathesan were introduced, separately and in combination, into the chromosome of Bacillus sphaericus 2297 by in vivo recombination. Two loci on the B. sphaericus chromosome were chosen as target sites for recombination: the binary toxin locus and the gene encoding the 36-kDa protease that may be responsible for the cleavage of the Mtx protein. Disruption of the protease gene did not increase the larvicidal activity of the recombinant strain against Aedes aegypti and Culex pipiens. Synthesis of the Cry11A and Cry11Ba toxins made the recombinant strains toxic to A. aegypti larvae to which the parental strain was not toxic. The strain containing Cry11Ba was more toxic than strains containing the added Cry11A or both Cry11A and Cry11Ba. The production of the two toxins together with the binary toxin did not significantly increase the toxicity of the recombinant strain to susceptible C. pipiens larvae. However, the production of Cry11A and/or Cry11Ba partially overcame the resistance of C. pipiens SPHAE and Culex quinquefasciatus GeoR to B. sphaericus strain 2297.
通过体内重组将来自苏云金芽孢杆菌以色列亚种的Cry11A和来自苏云金芽孢杆菌杰氏亚种的Cry11Ba分别及组合导入球形芽孢杆菌2297的染色体中。选择球形芽孢杆菌染色体上的两个位点作为重组的靶位点:二元毒素位点和编码可能负责Mtx蛋白裂解的36 kDa蛋白酶的基因。蛋白酶基因的破坏并未提高重组菌株对埃及伊蚊和致倦库蚊的杀幼虫活性。Cry11A和Cry11Ba毒素的合成使重组菌株对亲本菌株无毒的埃及伊蚊幼虫具有毒性。含有Cry11Ba的菌株比含有添加的Cry11A或同时含有Cry11A和Cry11Ba的菌株毒性更强。两种毒素与二元毒素一起产生并没有显著提高重组菌株对敏感致倦库蚊幼虫的毒性。然而,Cry11A和/或Cry11Ba的产生部分克服了致倦库蚊SPHAE和致倦库蚊GeoR对球形芽孢杆菌菌株2297的抗性。