Inducing Bacillus subtilis with Subtilin


Ncl uni logo.jpg

Newcastle University


Home Team Original Aims Software Modelling Proof of Concept Brick Wet Lab Conclusions

Home >> Wet Lab >> Protocols >> Inducing Bacillus subtilis with Subtilin

  • Make ON cultures of the transformed B. subtilis and B. subtilis ATCC6633 wild type
  • Add 50µl B. subtilis ON culture to 2mL LB.
  • Incubate for 1 hour at 37°C. (This is the optimum temperature for B. subtilis
  • Incubate for 3 hours at 30°C. (This is the optimum temperature for the reporter genes used (gfp and mCherry) as these are isolated from the cnidarians Aequorea victoria and Discosoma striata respectively. This enhances expression of the reporter genes.)
  • Divide the 2mL diluted culture into 2 screw-cap culture tubes and add subtilin to one (the other will be a control. To obtain subtilin, centrifuge the wild type ON culture and collect the supernatent. This should contain sectreted subtilin protein. Add this in the desired concentration (e.g for 1% induction, use 10µl supernatent in 1mL diluted culture.)
  • Incubate for 2 hours to allow induction of the insert and reporter gene.
  • Proceed to Preparing Bacillus subtilis for Microscopy