Difference between revisions of "Part:BBa K4907032"

 
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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
===Usage and Biology===
  
 
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<span class='h3bb'>Sequence and Features</span>
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===Biology===
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<i>ccdA</i>is another gene found within the <i>ccd>/i> operon, encoding the antidote protein (CcdA) that protects cells from the toxic effects of CcdB.
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===Usage and design===
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To verify the role of CcdA in resisting the CcdB toxin, we designed composite part: <partinfo>BBa_K4907138</partinfo> to characterize <i>ccdA</i>. The constructed circuit was transformed into <i>E. coli</i> DH10&beta;, followed by chloramphenicol selection of positive transformants, and confirmation was carried out through colony PCR and sequencing.
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<center><html><img src="https://static.igem.wiki/teams/4907/wiki/parts/yxy/parts/parts/i13453-b0034-ccda-b0015.png" width="400px"></html></center>
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<center><b>Fig. 1: Gene Circuit of BBa_K4907138</b></center>
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===Characterization===
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====Verification of dual-plasmid transformation====
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To validate the resistance of CcdA to CcdB, we performed a dual-plasmid transformation.
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Table. 1 Performance of different dual-plasmid systems in <i>E. coli</i> DH10&beta;
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The results showed that <i>E. coli</i> DH10&beta; transformed with both toxins and antitoxins also exhibited good growth. This confirmed the killing effect of the toxin CcdB again and the neutralization of antitoxin CcdA.
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===<span class='h3bb'>Sequence and Features</span>===
 
<partinfo>BBa_K4907032 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4907032 SequenceAndFeatures</partinfo>
  

Revision as of 18:04, 11 October 2023


ccdA

Biology

ccdAis another gene found within the ccd>/i> operon, encoding the antidote protein (CcdA) that protects cells from the toxic effects of CcdB.

Usage and design

To verify the role of CcdA in resisting the CcdB toxin, we designed composite part: BBa_K4907138 to characterize <i>ccdA. The constructed circuit was transformed into E. coli DH10β, followed by chloramphenicol selection of positive transformants, and confirmation was carried out through colony PCR and sequencing.

Fig. 1: Gene Circuit of BBa_K4907138

Characterization

Verification of dual-plasmid transformation

To validate the resistance of CcdA to CcdB, we performed a dual-plasmid transformation.


Table. 1 Performance of different dual-plasmid systems in E. coli DH10β The results showed that E. coli DH10β transformed with both toxins and antitoxins also exhibited good growth. This confirmed the killing effect of the toxin CcdB again and the neutralization of antitoxin CcdA.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]