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Proc Natl Acad Sci U S A. 1989 Jun;86(11):4037-41.

Location of the Bombyx mori specificity domain on a Bacillus thuringiensis delta-endotoxin protein.

Author information

1
Department of Biochemistry, Ohio State University, Columbus 43210.

Abstract

Bacillus thuringiensis produces different types of insecticidal crystal proteins (ICPs) or delta-endotoxins. In an effort to identify the insect specificity of ICP toxins, two icp genes were cloned into the Escherichia coli expression vector pKK223-3, and bioassays were performed with purified crystals. The type A protein [from an icpA1, or 4.5-kilobase (kb) gene, from B. thuringiensis var. kurstaki HD-1] was found to be 400 times more active against Bombyx mori than type C protein (from an icpC73, or 6.6-kb gene, from B. thuringiensis var. kurstaki HD-244). The type C protein was 9 times more active against Trichoplusia ni than the type A protein, while both have similar activity against Manduca sexta. To locate the specificity domain of the type A protein for B. mori, site-directed mutagenesis was used to introduce or remove restriction enzyme sites, facilitating the exchange of regions of the two genes. The hybrid genes were overexpressed, and purified ICP was used in bioassays. The B. mori specificity domain for the ICP A toxin is located in the amino-terminal portion of the hypervariable region between amino acids 332 and 450.

PMID:
2542961
PMCID:
PMC287383
DOI:
10.1073/pnas.86.11.4037
[Indexed for MEDLINE]
Free PMC Article

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