Efficient cloning and engineering of giant DNAs in a novel Bacillus subtilis genome vector.

@article{Itaya2000EfficientCA,
  title={Efficient cloning and engineering of giant DNAs in a novel Bacillus subtilis genome vector.},
  author={Mitsuhiro Itaya and Toshio Nagata and Toshihiko Shiroishi and Kyoko Fujita and Kenji Tsuge},
  journal={Journal of biochemistry},
  year={2000},
  volume={128 5},
  pages={869-75}
}
The Genome of Bacillus subtilis 168 was used for cloning and engineering of large-sized DNAs. A mouse genomic DNA of approximately 120 kb was cloned into a locus of the B. subtilis genome by ordered assembly of 20- to 50-kb mouse DNA segments. Cloned mouse DNA, maintained stably, was engineered through B. subtilis transformation and recombination. Creation of an I-PpoI recognition sequence at both ends of the insert facilitated its isolation by pulsed field gel electrophoresis. The basic… CONTINUE READING

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