Part:BBa_K1363201

Anti-LPS factor(LALF) regulated by IPTG

This part is a cell-kill system. Firstly,constitutive promoter could express enough Lacl that is useful for downstream. Secondly, the reporter in the middle of our part wouldn't show signal nor the cell be killed by the part behind unless IPTG or lactose added into the culture medium. No IPTG or lactose: GFP don't express ,G- won't be killed. IPTG or lactose exsisting : GFPs express ,G- will be dead.
G-:Gram negative bacteria

Results

We didn’t success in constructing this part. However, we directly complete the final circuit, which consists of lactose operator and signal peptide. Then we test it and it works well:

We used gradient plate method in our experiment. As we can see in the picture, the concentration of IPTG on the left of the medium is 0.1% while the concentration on the right is zero. We can see colony only on the right of the medium, the kill switch is effective which illustrates that IPTG can induce the expression of signal peptide and LALF so as to inhibit the growth of the E.coil. Also, a decrease of OD is observed too, after we added IPTG to the liquid culture medium, which means our kill switch did work well.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1208
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2085