title: Improved genetic transformation methods for the model alkaliphile Bacillus halodurans C-125.
abstract: OBJECTIVE
  Bacillus halodurans C-125 is a Gram-positive bacterium that was the first alkaliphilic species to have its genome completely sequenced. Despite its many years as a model for alkaliphily and source of industrially important enzymes, genetic manipulation of B. halodurans C-125 remains difficult, and therefore, we sought to develop a robust method to allow routine transformation of this organism.
  
  
  RESULTS
  A plasmid artificial modification system (PAM system, Yasui et al. 2008) for B. halodurans C-125 was created that increases transformation efficiency by 10- to 1000-fold. Also, recovering transformed protoplasts on succinate nutrient agar (SNA) yields faster, more robust colony recovery than on the traditional recovery medium. Combining these two techniques often allows recovery of transformants in as little as 48 h.
  
  
  CONCLUSIONS
  Use of the B. halodurans C-125 PAM system and SNA greatly improves the efficiency and speed of protoplast transformation of B. halodurans C-125.
  
  
  CONCLUSIONS
  These techniques allow routine genetic manipulation of B. halodurans C-125, a model alkaliphilic bacterium with important industrial properties.
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, USA.
authors.forename: J G
authors.initials: JG
authors.lastname: Wallace
authors.forename: R R
authors.initials: RR
authors.lastname: Breaker
mesh_terms: D001407:Bacillus / Q000235:genetics*; D005821:Genetic Techniques; D008954:Models, Biological; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic