Difference between revisions of "Part:BBa K2922028"

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===Summary===
 
===Summary===
 
Abbreviated as Nimm, the molecular weight of Colicin-N immunity protein is 15 kDa protein, and the protein is encoded by gene  
 
Abbreviated as Nimm, the molecular weight of Colicin-N immunity protein is 15 kDa protein, and the protein is encoded by gene  
<i>cni</i>. In wild strains of <i>E.coli</i> ,the protein is able to protect cells, which harbors the plasmid ColN encoding Colicin-N, against Colicin-N.[1] This protein will prevent <i>E.coli</i> secreting Colicin-N from killing themselves.[2] (Fig.1)
+
<i>cni</i>. In wild strains of <i>E.coli</i>,the protein is able to protect cells, which harbors the plasmid ColN encoding Colicin-N, against Colicin-N.[1]This protein will prevent <i>E.coli</i> secreting Colicin-N from killing themselves.[2] (Fig.1)
 
<table><tr><th>[[File:Ndesign.png|thumb|300px|Fig.1 Nimm prevents <i>E.coli</i> BL21 (DE3) secreting Colicin-E1 from killing themselves.]]</th><th></table>
 
<table><tr><th>[[File:Ndesign.png|thumb|300px|Fig.1 Nimm prevents <i>E.coli</i> BL21 (DE3) secreting Colicin-E1 from killing themselves.]]</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 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.
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 getting the target separate fragment- 396bp. (Fig.2)
+
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-396bp. (Fig.2)
 
<table><tr><th>[[File:Eimm Nimm.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:Eimm Nimm.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>
 
===Reference===
 
===Reference===

Revision as of 14:21, 19 October 2019


Colicin-N immunity protein coding region

Summary

Abbreviated as Nimm, the molecular weight of Colicin-N immunity protein is 15 kDa protein, and the protein is encoded by gene cni. In wild strains of E.coli,the protein is able to protect cells, which harbors the plasmid ColN encoding Colicin-N, against Colicin-N.[1]This protein will prevent E.coli secreting Colicin-N from killing themselves.[2] (Fig.1)

Fig.1 Nimm prevents E.coli BL21 (DE3) secreting Colicin-E1 from killing themselves.

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. After new molecular cloning experiments, we do Enzyme-Cut identification to certify the plasmid is correct. We use the XbaI and PstI to cut the plasmid, then we get the target separate fragment-396bp. (Fig.2)

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

Reference

[1] A. P. Pugsley, The immunity and lysis genes of ColN plasmid pCHAP4. Molecular & General Genetics Mgg 211, 335-341 (1988).
[2] 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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]