Studies on the Genome Engineering Using Bacillus subtilis 168.

Genome engineering using a synthetic gene circuit in Bacillus subtilis

Genome engineering without leaving foreign DNA behind requires an efficient counter-selectable marker system. Here, we developed a genome engineering method in Bacillus subtilis using a synthetic gene circuit as a counter-selectable marker system. The system contained two repressible promoters (B. subtilis xylA (Pxyl) and spac (Pspac)) and two repressor genes (lacI and xylR). Pxyl-lacI was inte...

Suppressor system in Bacillus subtilis 168.

Multiple auxotrophic strains of Bacillus subtilis 168 were tested for joint one-step reversion of two or more auxotrophic markers to the wild-type phenotype. Mu8u5u5, a strain requiring leucine, methionine, and threonine, yielded revertants that grew without added methionine or threonine and proved to have a suppressor gene. When transferred by transformation with deoxyribonucleic acid, this su...

Bacillus subtilis 168 4 5 6

Restriction on conjugational transfer of pLS20 in Bacillus subtilis 168.

Conjugational transfer of pLS20 in Bacillus subtilis Marburg 168 is restricted by the BsuM restriction-modification system. Restriction efficiency was measured using pLS20 derivatives possessing various numbers of XhoI sites, which are known to be recognized by BsuM. An increase in XhoI sites clearly reduced the conjugational efficiency of pLS20 as compared with that of pUB110 plasmid lacking X...

Region dependent efficiency for recombinational transfer of the Bacillus subtilis 168 genome.

Submega-sized regions of the Bacillus subtilis genome were cloned to plasmid by the B. subtilis Recombinational Transfer (BReT) method. BReT efficiency depends not only on the genome location but also on the choice of sequences for simultaneous homologous recombination during BReT. In an extreme case, a 91-kb region that was unsuccessful on the first attempt was obtained when the slightly shift...