New integrative method to generate Bacillus subtilis recombinant strains free of selection markers.

@article{Brans2004NewIM,
  title={New integrative method to generate Bacillus subtilis recombinant strains free of selection markers.},
  author={Alain Brans and Patrice Fil{\'e}e and Andy Chevign{\'e} and Aurore Claessens and Bernard Joris},
  journal={Applied and environmental microbiology},
  year={2004},
  volume={70 12},
  pages={7241-50}
}
The novel method described in this paper combines the use of blaI, which encodes a repressor involved in Bacillus licheniformis BlaP beta-lactamase regulation, an antibiotic resistance gene, and a B. subtilis strain (BS1541) that is conditionally auxotrophic for lysine. We constructed a BlaI cassette containing blaI and the spectinomycin resistance genes and two short direct repeat DNA sequences, one at each extremity of the cassette. The BS1541 strain was obtained by replacing the B. subtilis… CONTINUE READING