Difference between revisions of "Part:BBa K1976054"
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         <h1>Colicin E2 DNase with B0034 and T7 promoter</h1>
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         <h1>Colicin E2 DNase and T7 promoter</h1>
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Colicin E2 is a peptide, that is toxic to coliform bacteria. It is one of a group of proteins of this type, it being a nicking endonuclease. It has a corresponding activity inhibiting protein called immunity protein Im2, which is also in the registry under the name.<br>
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This part encodes the DNase fragment of colicin E2 with the RBS B0034 (part <a href="https://parts.igem.org/Part:BBa_K1976048">BBa_K1976048</a>) and a T7 promoter.
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This part can be created by digesting Colicin E2 with Trypsin.
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The Colicin E2 immunity protein (<a href="https://parts.igem.org/Part:BBa_K1976027">BBa_K1976027</a> and <a href="https://parts.igem.org/Part:BBa_K1976028">BBa_K1976028</a>) can be used in combination with the Colicin E2 DNase domain, or this part as an inhibitor. Since it binds very tightly to the immunity protein, it can also be used in fusing proteins to one another.
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The colicin DNase fragment is an active DNase and can be used for DNA degradation. The T7 promoter is an inducable promoter wich ensures expression of the so called miniColicin after induction with IPTG in cells which have a T7 promoter system.
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===<h2>Characteristics</h2>===
 
===<h2>Characteristics</h2>===
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The DNase fragment of colicin E2 has a molecular weight of 15.2 kDa and consists of 133 residues what conforms to about one third to colicin. We used only one of the three domains of colicin since the metabolic burdon would be significant smaller. The activity tests showed that the so called miniColicin has functional DNase activity.
  
 
<h2>Sequence and Features</h2>
 
<h2>Sequence and Features</h2>
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<partinfo>BBA_K1976049 SequenceAndFeatures</partinfo>
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<partinfo>BBA_K1976054 SequenceAndFeatures</partinfo>
 
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Latest revision as of 03:26, 28 October 2016

Colicin E2 DNase and T7 promoter

This part encodes the DNase fragment of colicin E2 with the RBS B0034 (part BBa_K1976048) and a T7 promoter.

Figure 1:Figure 5: growth of BL21 E. coli transformed with a): BBa_K1976054 and b): BBa_K1976055.


Usage

The colicin DNase fragment is an active DNase and can be used for DNA degradation. The T7 promoter is an inducable promoter wich ensures expression of the so called miniColicin after induction with IPTG in cells which have a T7 promoter system.

Characteristics

The DNase fragment of colicin E2 has a molecular weight of 15.2 kDa and consists of 133 residues what conforms to about one third to colicin. We used only one of the three domains of colicin since the metabolic burdon would be significant smaller. The activity tests showed that the so called miniColicin has functional DNase activity.

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 66
  • 1000
    COMPATIBLE WITH RFC[1000]


References
[1] Cascales et al, Colicin Biology, Microbiology and Molecular Biology Reviews, vol. 71, pp. 158-229, 2007