Difference between revisions of "Part:BBa K4907032"

Revision as of 18:05, 11 October 2023

ccdA

Biology

ccdAis another gene found within the ccd 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 a composite part: BBa_K4907138 to characterize 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]

@@ Line 7: / Line 7: @@
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 ===Biology===
-<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.
+<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.
 ===Usage and design===
-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.
+To verify the role of CcdA in resisting the CcdB toxin, we designed a 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.
 <center><html><img src="https://static.igem.wiki/teams/4907/wiki/parts/yxy/parts/parts/i13453-b0034-ccda-b0015.png" width="400px"></html></center>
 <center><b>Fig. 1: Gene Circuit of BBa_K4907138</b></center>