Part:BBa_K302035

Sucrose-limitation induced kill switch

Encodes a stable non-specific ribonuclease toxin (mazF) and its inhibitory antitoxin (mazE) in Bacillus subtilis. Contains Sucrose sensitive inducer that will only allow coding sequence translation in the presence of sucrose. These genes are used in Bacillus subtilis to provide a toxin-antitoxin kill switch in various stressful conditions. When translation of both genes is turned off by sucrose limitation mazE will be degraded faster than mazF. There is then no inhibiton of mazF, killing the cell.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 363
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

User Reviews

james_zhang

In 2017 SZU-China iGEM team has characterized the output of this part in B.subtilis WB800. The result was documented in the experience page and the main page of BBa_K2232025

iGEM2017 SZU-China confirmed the function of BBa_K302035.

We made characterization of this part in our chassis B.subtilis WB800.The sucrose was added in the culture medium when the value of OD₆₀₀ reach stationary phase initially, and the final concentration of sucrose was 20mmol/L. Then we recorded the variation of OD₆₀₀ by interval of 4 hours and the result is shown as follow.

Fig. The OD₆₀₀ of original strain WB800 and recombinant B.subtilis WB800_MazEF recorded by interval of 4 hours.The red arrow represents the adjunction of Sucrose (20mmol/L).

As can be seen from the figure above, the Sucrose-limitation induced kill switch in B.subtilis didn’t show obvious function on our chassis.The reasons may be the concentration of sucrose wasn’t suitable or the degradation of ribonuclease toxin (mazF) is too fast.