Part:BBa_K2752000

Part3

The use of BBa_K2752000 in combination with BBa_K2752001 prevents the loss of the plasmids.Reducing the level of uric acid in the environment can cause the lytic enzyme to function, resulting in the death of the cells.

Sequence and Features

Characterization

HucO is regulated by HucR. When uric acid is abscent, HucR can bind to hucO making downstream gene silencing. Lysis1 and Lysis2 are lyases. Lysis1siRNA286 can inhibit Lysis1, and Lysis2siRNA106 can inhibit Lysis2. When both plasmids are simultaneously expressed, the lyases are inhibited. But if any plasmid is lost or HucR combined hucO，part3 will not be expressed. And the bacterium is cleaved by the Lysis2 . A series of things happens when bacteria are excreted: First, the concentration of uric acid get lower. Second, the hucO of part3 is suppressed by HucR and cannot work. Next, Lysis2 siRNA106 cannot inhibit Lysis2. Finally, the bacteria are cleaved by Lysis2. To test the lethal ability of part2 and part3, and the regulation of uric acid on part3 we did four sets of experiments.

Figure 1. Plasmid map of part2 and part3。

Figure 2. Colony number regulation experiment result.Blank is a plasmid-free BL21 strain. Part3 is a BL21 strain containing the part3 plasmid. Part2 is a BL21 strain containing the part2 plasmid. Part3-with uric acid is a BL21 strain containing part3 plasmid cultured with a uric acid （The final concentration of uric acid was adjusted to 4×10-4M.）

Assembly Compatibility:

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

  Illegal XhoI site found at 568
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1257
  Illegal AgeI site found at 1327
  Illegal AgeI site found at 1465
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 304
  Illegal SapI.rc site found at 268