Coding

Part:BBa_K2913021

Designed by: Yannan Wang   Group: iGEM19_NEFU_China   (2019-10-20)
Revision as of 10:04, 20 October 2019 by LLLLLLYW (Talk | contribs) (result)


Cytosine deaminase

An enzyme that oxidizes 5-fluorouracil to 5-fluorouracil

Usage and Biology

result

Cytosine deaminase (CD) is the enzyme that can transform cytosine into uracil. CD’s function is to convert nontoxic 5-Flucytosine (5-FC) into toxic 5-fluorouracil 5-FU, which has an extreme damage to the bacteria.

We added gradient of 5-FC culturing the bacteria and monitor their growth.

Fig.1 Growth of E.coli at different concentration of 5-FC. Foster the bacteria (keeping the quantity of the bacteria same before 1% passaging : 2μl bacteria, OD=3.5) with a concentration gradient of 5-FC in the LB mediacontaining 50µg/ml Amp and str): 500mg/l, 250mg/l, 100mg/l, 50mg/l, 25mg/l, 0mg/l. A. The bacteria (E.coli) using T7 to express CD. B. The bacteria (E.coli) can`t express CD.


As a positive control, we added gradient of 5-FU culturing the bacteria and monitor their growth.

Fig.2 Growth of bacteria at different concentrations of 5-FU. Foster the bacteria (keeping the quantity of the bacteria same before 1% passaging: 2ul bacteria, OD=3.5) with a concentration gradient of 5-FC in the LB media (containing 50 µg/ml Amp and str): 500mg/l, 250mg/l, 100mg/l, 50mg/l, 25mg/l, 0mg/l. A. The bacteria using T7 to express CD. B. The original bacteria that without the CD expression plasmid.

For more details about the part, please visit NEFU_China 2019 wiki.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1140
  • 1000
    COMPATIBLE WITH RFC[1000]


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