Difference between revisions of "Part:BBa K2922024"

(Identification)
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As a kind of bacteriocin secreted by <i>E.coli</i>, colicin can kill other related bacteria that can’t secret specific immunity proteins.  
 
As a kind of bacteriocin secreted by <i>E.coli</i>, colicin can kill other related bacteria that can’t secret specific immunity proteins.  
 
For Colicin-E1, it is a type of pore-forming colicin encoded by gene <i>cea</i>.This class of transmembrane toxins depolarize the cytoplasmic membrane, leading to dissipation of cellular energy.[1] (Fig.1)
 
For Colicin-E1, it is a type of pore-forming colicin encoded by gene <i>cea</i>.This class of transmembrane toxins depolarize the cytoplasmic membrane, leading to dissipation of cellular energy.[1] (Fig.1)
<table><tr><th>[[File:E1design.png|thumb|300px|Fig.1 Colicin-E1 can kill other bacteria.]]</th><th></table>
+
<table><tr><th>[[File:E1design.png|thumb|720px|Fig.1 Colicin-E1 can kill other bacteria.]]</th><th></table>
 
===Identification===
 
===Identification===
When we are building this circuit, we are doing the nucleic acid gel electrophoresis experiment to verify. After the circuit is built, we send the plasmid to sequence, and get the correct sequencing.
+
When we were building this circuit, we did the nucleic acid gel electrophoresis experiment to verify. After the circuit was built, we sent the plasmid to sequence, and got the correct sequencing.
After new molecular cloning experiments, we do Enzyme-Cut identification to certify the plasmid is correct. We use the <i>Xba</i>I and <i>Pst</i>I to cut the plasmid, then we get the target separate fragment-1569bp. (Fig.2)
+
After new molecular cloning experiments, we did Enzyme-Cut identification to certify the plasmid was correct. We used the <i>Xba</i>I and <i>Pst</i>I to cut the plasmid, then we got the target separate fragment-1569bp. (Fig.2)
 
<table><tr><th>[[File:E1 N.png|thumb|300px|Fig.2 The result of this plasmid cut with enzyme <i>Xba</i>I and <i>Pst</i>I.]]</th><th></table>
 
<table><tr><th>[[File:E1 N.png|thumb|300px|Fig.2 The result of this plasmid cut with enzyme <i>Xba</i>I and <i>Pst</i>I.]]</th><th></table>
  

Revision as of 03:03, 20 October 2019


Colicin-E1 coding region

Summary

As a kind of bacteriocin secreted by E.coli, colicin can kill other related bacteria that can’t secret specific immunity proteins. For Colicin-E1, it is a type of pore-forming colicin encoded by gene cea.This class of transmembrane toxins depolarize the cytoplasmic membrane, leading to dissipation of cellular energy.[1] (Fig.1)

Fig.1 Colicin-E1 can kill other bacteria.

Identification

When we were building this circuit, we did the nucleic acid gel electrophoresis experiment to verify. After the circuit was built, we sent the plasmid to sequence, and got the correct sequencing. After new molecular cloning experiments, we did Enzyme-Cut identification to certify the plasmid was correct. We used the XbaI and PstI to cut the plasmid, then we got the target separate fragment-1569bp. (Fig.2)

Fig.2 The result of this plasmid cut with enzyme XbaI and PstI.

Reference

[1] E. Cascales et al., Colicin biology. Microbiol Mol Biol Rev 71, 158-229 (2007).

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 AgeI site found at 1312
    Illegal AgeI site found at 1369
    Illegal AgeI site found at 1501
  • 1000
    COMPATIBLE WITH RFC[1000]