Part:BBa_K649302

rocF-Arg box

rocF is cording region of Arginase. ArgBox is operator sequence of ArgI(E. coli).

These were obtaind by colony-PCR, and ligated into pTrc99A. rocF had 2 EcoRI sites, so we changed these sites on pTrc99A.

At last, backbone was changed to pSB1C3.

To characterize this part, we constructed the composite part(BBa_K649402).

Urea concentration detected in bacterial samples on pSB3K3
This work is done by Natsuki Kubo.

Introduction of BBa_K649402 containing arginase coding gene (rocF) and Arg Box (an arg operator capable of binding the arginine repressor) led to production of some urea in strain MG1655 (argR +) and strain JE6852 (argR -). But in both strains, the amount of urea produced was not much different from that when BBa_K649301 containing rocF but not Arg box was introduced. This result shows that rocF is working but Arg box is not working efficiently in this part. The reason why Arg box is not working is probably that low-copy-number plasmid used here is not capable of introducing enough number of Arg boxes to effectively deactivate the arginine repressor.

For more information, see [http://2011.igem.org/Team:Tokyo_Tech/Projects/Urea-cooler/data#2. our work in Tokyo_Tech 2011 wiki.]

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 137
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]