Cloning and Characterization of a Cytolytic and Mosquito Larvicidal δ-Endotoxin from Bacillus thuringiensis subsp. darmstadiensis
| Autor: | Sutipa Tanapongpipat, Plearnpis Luxananil, Namchai Chewawiwat, Boonhiang Promdonkoy, Sakol Panyim |
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| Rok vydání: | 2003 |
| Předmět: |
Sequence analysis
Bacterial Toxins Molecular Sequence Data Bacillus thuringiensis Aedes aegypti Molecular cloning medicine.disease_cause Hemolysis Applied Microbiology and Biotechnology Microbiology Hemolysin Proteins Bacterial Proteins Aedes Escherichia coli medicine Animals Amino Acid Sequence Cloning Molecular Peptide sequence Bacillus thuringiensis Toxins Base Sequence biology fungi Sequence Analysis DNA General Medicine biology.organism_classification Proteinase K Bacillales Molecular biology Endotoxins Culex Larva biology.protein Endopeptidase K |
| Zdroj: | Current Microbiology. 46:94-98 |
| ISSN: | 1432-0991 0343-8651 |
| DOI: | 10.1007/s00284-002-3823-5 |
| Popis: | The cytolytic delta-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 m M Na(2)CO(3), 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 end- point 0.25 microgram/ml) and was toxic to Culex quinquefasciatus and Aedes aegypti larvae (LC(50) 0.5-1.0 microgram/ml). |
| Databáze: | OpenAIRE |
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