Publication: Cloning and characterization of a cytolytic and mosquito larvicidal δ-endotoxin from Bacillus thuringiensis subsp. darmstadiensis
Issued Date
2003-02-01
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ISSN
03438651
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2-s2.0-0038192410
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Mahidol University
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SCOPUS
Bibliographic Citation
Current Microbiology. Vol.46, No.2 (2003), 94-98
Suggested Citation
Boonhiang Promdonkoy, Namchai Chewawiwat, Sutipa Tanapongpipat, Plearnpis Luxananil, Sakol Panyim Cloning and characterization of a cytolytic and mosquito larvicidal δ-endotoxin from Bacillus thuringiensis subsp. darmstadiensis. Current Microbiology. Vol.46, No.2 (2003), 94-98. doi:10.1007/s00284-002-3823-5 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/20929
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Title
Cloning and characterization of a cytolytic and mosquito larvicidal δ-endotoxin from Bacillus thuringiensis subsp. darmstadiensis
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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).