Difference between revisions of "Part:BBa K2913021"
 
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<partinfo>BBa_K2913021 short</partinfo>
 
<partinfo>BBa_K2913021 short</partinfo>
  
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An enzyme that oxidizes 5-fluorouracil to 5-fluorouracil
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An enzyme that oxidizes 5-flucytosine to 5-fluorouracil
  
 
===Usage and Biology===
 
===Usage and Biology===
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=result=
 
=result=
  
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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.
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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.
  
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We added gradient of 5-FC culturing the bacteria and monitor their growth.
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We added gradient of 5-FC culturing the bacteria and monitored their growth.
  
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[[Image:T--NEFU_China--parts--CD1.png|600px|thumb|left|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.]]
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[[Image:T--NEFU_China--parts--CD1.png|600px|thumb|left|Fig.1 Growth of <i>E.coli</i> at different concentration of 5-FC. Fostered the bacteria (keeping the quantity of the bacteria same before 1% passaging : 2 μl bacteria, OD=3.5) with a concentration gradient of 5-FC in the LB media (containing 50µg/ml Amp and Str): 500 mg/l, 250 mg/l, 100 mg/l, 50 mg/l, 25 mg/l, 0 mg/l. A. The bacteria (<i>E.coli</i>) using T7 to express CD. B. The bacteria (<i>E.coli</i>) can`t express CD.]]
  
  
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<p style="margin-top:24em;">As a positive control, we added gradient of 5-FU culturing the bacteria and monitor their growth.</p>
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<p style="margin-top:24em;">As a positive control, we added gradient of 5-FU culturing the bacteria and monitored their growth.</p>
  
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[[File:T--NEFU_China--parts--CD2.png|600px|thumb|left|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.]]
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[[File:T--NEFU_China--parts--CD2.png|600px|thumb|left|Fig.2 Growth of bacteria at different concentrations of 5-FU. Fostered the bacteria (keeping the quantity of the bacteria same before 1% passaging: 2 μl bacteria, OD=3.5) with a concentration gradient of 5-FC in the LB media (containing 50 µg/ml Amp and Str): 500 mg/l, 250 mg/l, 100 mg/l, 50 mg/l, 25 mg/l, 0 mg/l. A. The bacteria using T7 to express CD. B. The original bacteria  that without the CD expression plasmid.]]
  
 
<p style="margin-top:26em;">For more details about the part, please visit NEFU_China 2019 wiki.</p>
 
<p style="margin-top:26em;">For more details about the part, please visit NEFU_China 2019 wiki.</p>
  
 
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<p style="margin-top:26em;"><span class='h3bb'>Sequence and Features</span></p>
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<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2913021 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2913021 SequenceAndFeatures</partinfo>
  

Latest revision as of 17:10, 21 October 2019


Cytosine deaminase

An enzyme that oxidizes 5-flucytosine 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 monitored their growth.

Fig.1 Growth of E.coli at different concentration of 5-FC. Fostered the bacteria (keeping the quantity of the bacteria same before 1% passaging : 2 μl bacteria, OD=3.5) with a concentration gradient of 5-FC in the LB media (containing 50µg/ml Amp and Str): 500 mg/l, 250 mg/l, 100 mg/l, 50 mg/l, 25 mg/l, 0 mg/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 monitored their growth.

Fig.2 Growth of bacteria at different concentrations of 5-FU. Fostered the bacteria (keeping the quantity of the bacteria same before 1% passaging: 2 μl bacteria, OD=3.5) with a concentration gradient of 5-FC in the LB media (containing 50 µg/ml Amp and Str): 500 mg/l, 250 mg/l, 100 mg/l, 50 mg/l, 25 mg/l, 0 mg/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]