Difference between revisions of "Part:BBa K1976054"
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         <h1>Colicin E2 DNase with B0034 and T7 promoter</h1>
 
         <h1>Colicin E2 DNase with B0034 and T7 promoter</h1>
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Generator for the Colicin DNase domain <a href="https://parts.igem.org/Part:BBa_K1976048">BBa_K1976048</a>.
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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 exhibited signs of leakage, that can kill the host if cells are transformed with it, that have a T7 promoter system, due to the endonuclease activity of the Colicin E2 DNase domain.  
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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 comform 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>

Revision as of 02:54, 28 October 2016

Colicin E2 DNase with B0034 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 comform 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