A simple method for separation of the protein crystal from Bacillus thuringiensis using carboxymethyl cellulose column chromatography
| Autor: | Roop Singh Bora, Vaithilingam Sekar, G. Srinivas, M. G. Murty |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Microbiology (medical)
Gel electrophoresis Chromatography biology Sodium fungi chemistry.chemical_element biology.organism_classification Microbiology Bacillales Carboxymethyl cellulose chemistry.chemical_compound Column chromatography chemistry Bacillus thuringiensis medicine Protein crystallization Molecular Biology Sodium acetate medicine.drug |
| Zdroj: | Journal of Microbiological Methods. 19:103-110 |
| ISSN: | 0167-7012 |
| DOI: | 10.1016/0167-7012(94)90041-8 |
| Popis: | A simple and a rapid technique to separate intact crystals from spores and cell debris of Bacillus thuringiensis using a carboxymethyl cellulose (CM-cellulose) column is described. B. thuringiensis subsp. kurstaki, a toxic strain to lepidopteran insects that produces small bipyramidal crystals, and B. thuringiensis subsp. yunnanensis, a nontoxic strain producing unusually large bipyramidal and spindle-shaped crystals were used as test organisms. Spores and cell debris were washed in sodium acetate and sodium phosphate buffers and the crystals bound to the CM-cellulose were eluted with Tris-EDTA buffer, regardless of their size and shape. Light microscopy and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the purity of the crystal was > 98%. The percent recoveries of crystals from B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. yunnanensis were approximately 14 and 25, respectively. |
| Databáze: | OpenAIRE |
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