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Title: Cloning and characterization of a cytolytic and mosquito larvicidal δ-endotoxin from Bacillus thuringiensis subsp. darmstadiensis
Authors: Boonhiang Promdonkoy
Namchai Chewawiwat
Sutipa Tanapongpipat
Plearnpis Luxananil
Sakol Panyim
Thailand National Center for Genetic Engineering and Biotechnology
Mahidol University
Keywords: Immunology and Microbiology
Issue Date: 1-Feb-2003
Citation: Current Microbiology. Vol.46, No.2 (2003), 94-98
Abstract: The cytolytic δ-endotoxin gene from Bacillus thuringiensis subsp. darmstadiensis was amplified from genomic DNA by PCR by using primers designed from the sequence of cyt2Aa1 gene of B. thuringiensis subsp. kyushuensis. DNA sequence analysis revealed an open reading frame translating to a 259-amino acid sequence. The cloned gene was designated cyt2Aa2. This gene was highly expressed in Escherichia coli as inclusion bodies that could be solubilized in 50 mM Na2CO3, pH 10.5. Activation of the solubilized protoxin by proteinase K (1% wt/wt, proteinase K/protoxin) yielded the active fragment of about 23 kDa. Cyt2Aa2 showed high hemolytic activity against sheep erythrocytes (hemolytic endpoint 0.25 μg/ml) and was toxic to Culex quinquefasciatus and Aedes aegypti larvae (LC50 0.5-1.0 μg/ml).
ISSN: 03438651
Appears in Collections:Scopus 2001-2005

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