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| Content Provider | Springer Nature Link |
|---|---|
| Author | Vojcic, Ljubica Despotovic, Dragana Martinez, Ronny Maurer, Karl Heinz Schwaneberg, Ulrich |
| Copyright Year | 2012 |
| Abstract | Bacillus subtilis strains are used for extracellular expression of enzymes (i.e., proteases, lipases, and cellulases) which are often engineered by directed evolution for industrial applications. B. subtilis DB104 represents an attractive directed evolution host since it has a low proteolytic activity and efficient secretion. B. subtilis DB104 is hampered like many other Bacillus strains by insufficient transformation efficiencies (≤103 transformants/μg DNA). After investigating five physical and chemical transformation protocols, a novel natural competent transformation protocol was established for B. subtilis DB104 by optimizing growth conditions and histidine concentration during competence development, implementing an additional incubation step in the competence development phase and a recovery step during the transformation procedure. In addition, the influence of the amount and size of the transformed plasmid DNA on transformation efficiency was investigated. The natural competence protocol is “easy” in handling and allows for the first time to generate large libraries (1.5 × 105 transformants/μg plasmid DNA) in B. subtilis DB104 without requiring microgram amounts of DNA. |
| Starting Page | 487 |
| Ending Page | 493 |
| Page Count | 7 |
| File Format | |
| ISSN | 01757598 |
| Journal | Applied Microbiology and Biotechnology |
| Volume Number | 94 |
| Issue Number | 2 |
| e-ISSN | 14320614 |
| Language | English |
| Publisher | Springer-Verlag |
| Publisher Date | 2012-03-07 |
| Publisher Place | Berlin, Heidelberg |
| Access Restriction | Subscribed |
| Subject Keyword | B. subtilis DB104 Directed evolution Natural competence Transformation protocol Microbiology Biotechnology Microbial Genetics and Genomics |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Applied Microbiology and Biotechnology Biotechnology |
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