Sequential insertion of multiple I-SceI recognition sites at designed loci of the Bacillus subtilis 168 genome.
Bacillus subtilis 168 is the only bacterium-based host serving for the cloning of giant DNA above 1.000 kbp. As rapid verification of the genome structure is crucial during the cloning process, six of 18-base sequence recognized by endonuclease I-SceI were sequentially created in the B. subtilis 168 genome. The established method and materials should be of use for other B. subtilis derivatives.
Mitsubishi Kagaku Institute of Life Sciences, Machida, Tokyo, Japan. mita2001@sfc.keio.ac.jp
Mitsuhiro
M
Itaya
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics*; D001665:Binding Sites; D004720:Endonucleases / Q000378:metabolism*; D056426:Genetic Loci / Q000235:genetics*; D016680:Genome, Bacterial / Q000235:genetics*; D016254:Mutagenesis, Insertional
D004720:Endonucleases