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  
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<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)
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<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.<ref>[1]https://link.springer.com/article/10.1007/BF00330613</ref>
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<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>
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This protein will prevent <i>E.coli</i> secreting Colicin-N from killing themselves.<ref>[2]https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1847374/</ref>(Fig.1)
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<table><tr><th>[[File:Ndesign.png|thumb|720px|Fig.1 Nimm prevents <i>E.coli</i> BL21 (DE3) secreting Colicin-E1 from killing themselves.]]</th><th></table>
 
===Identification===
 
===Identification===
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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.
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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.
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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)
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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-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>
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===Reference===
 
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[1] A. P. Pugsley, The immunity and lysis genes of ColN plasmid pCHAP4. Molecular & General Genetics Mgg 211, 335-341 (1988).
 
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<br>
 
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[2] E. Cascales et al., Colicin biology. Microbiol Mol Biol Rev 71, 158-229 (2007).
 
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<br>
 
  
 
===Sequence and Features===
 
===Sequence and Features===
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<partinfo>BBa_K2922028 parameters</partinfo>
 
<partinfo>BBa_K2922028 parameters</partinfo>
 
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===Reference===

Latest revision as of 17:58, 20 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 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-396bp. (Fig.2)

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

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]


Reference

  1. ↑ [1]https://link.springer.com/article/10.1007/BF00330613
  2. ↑ [2]https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1847374/