We attempted to use Bacillus subtilis phage phi 1 as a gene-cloning vector since the phi 1 genome was found to have few cleavage sites upon digestion with several kinds of restriction endonucleases. A phi 1 stock supplied by J. Ito (University of Arizona, Tucson, USA) consisted of two phages, phi 1E
A Useful Cloning Vector for Bacillus subtilis
✍ Scribed by Børge Diderichsen; Gitte Bak Poulsen; Steen T. Jørgensen
- Book ID
- 112263162
- Publisher
- Elsevier Science
- Year
- 1993
- Tongue
- English
- Weight
- 147 KB
- Volume
- 30
- Category
- Article
- ISSN
- 0147-619X
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Cloning in Escherichia coli and Bacillus subtilis was carried out using the bifunctional plasmid pDH5060. B. subtilis chromosomal DNA and pDH5060 DNA were digested with either BamHI or SalI, then annealed, ligated, and transformed into E. coli SK2267. Transformants containing sequences ligated into
Recombinant plasmid DNA cloned in E. coli via the bifunctional vector pDH5060 suffered deletions when returned to B. subtilis. However, DNA preparations of identical chimeras containing homologous or heterologous sequences stably transformed B. subtilis at high efficiency when isolated from B. subti