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The fragment containing the gene encoding the cytolytic Cyt1Ab1 protein fromBacillus thuringiensis subsp.medellin and its flanking sequences (I. Thiery, A. Delécluse, M. C. Tamayo, and S. Orduz, Appl. Environ. Microbiol. 63:468–473, 1997) was introduced intoBacillus sphaericus toxic strains 2362, 2297, and Iab872 by electroporation with the shuttle vector pMK3. Only small amounts of the protein were produced in recombinant strains 2362 and Iab872. The protein was detected in these strains only by Western blotting and immunodetection with antibody raised against Cyt1Ab1 protein. Large amounts of Cyt1Ab1 protein were produced inB. sphaericus recombinant strain 2297, and there was an additional crystal, other than that of the binary toxin, within the exosporium. The production of the Cyt1Ab1 protein in addition to the binary toxin did not increase the larvicidal activity of theB. sphaericus recombinant strain against susceptible mosquito populations ofCulex pipiens orAedes aegypti . However, it partially restored (10 to 20 times) susceptibility of the resistant mosquito populations ofC. pipiens (SPHAE) andCulex quinquefasciatus (GeoR) to the binary toxin. The Cyt1Ab1 protein produced in recombinantB. thuringiensis SPL407(pcyt1Ab1 ) was synthesized in two types of crystal—one round and with various dense areas, surrounded by an envelope, and the other a regular cuboid crystal, very similar to that found in theB. sphaericus recombinant strain.

applied and environmental microbiology

The Introduction into Bacillus sphaericus of the Bacillus thuringiensis subsp. medellin cyt1Ab1 Gene Results in Higher Susceptibility of Resistant Mosquito Larva Populations to B. sphaericus