the best growing condition for Bacillus subtilis WB800N

To ensure the expression of our T-vaccine, we do some experiments to find out the best growing condition for Bacillus subtilis WB800N, we finally choose five independent factors: temperature, inoculation time, peptone, yeast extract and NaCl. The following table displays their levels:

图无法导出 Roughly, we could consider the treatment of No. 15 medium (Temperature 35℃, Time 12h, Peptone 15, Yeast Extract 7.5, NaCl 15)as the maximal condition for Bacillus subtilis.

Modeling on the suicide mechanism of the reporter system

To eliminate potential safety problem, we constructed a suicide system on engineered bacteria to ensure biosafety. As the only one loaded with kill switch, the engineered reporter bacteria is responsible for eliminating all siblings in T-vaccine. Killing is mediated by the exported toxic protein SdpC. Extracellular SdpC induces the synthesis of an immunity protein, SdpI, which protects toxin-producing cells from being killed. SdpI, a polytopic membrane protein, is encoded by a two-gene operon under sporulation control that contains the gene for an autorepressor, SdpR.