Difference between revisions of "Part:BBa K1976055"
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         <h1>trypsin fragment of Colicin E2 with B0034 and T7 promoter</h1>
 
         <h1>trypsin fragment of Colicin E2 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 trypsin 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.
 
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===<h2>Characteristics</h2>===
 
===<h2>Characteristics</h2>===
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The DNase trypsin fragment of colicin E2 consists of 137 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. This fragment differs to the part 50 in 5 additional amino acids since a trypsin digestion described in the literature leaded to this fragment. So we wanted to check out any difference in the activities between the DNase fragment with and without theese additional amino acids. The activity tests showed that the so called miniColicin has functional DNase activity.
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<h2>Sequence and Features</h2>
 
<h2>Sequence and Features</h2>
 
<partinfo>BBA_K1976055 SequenceAndFeatures</partinfo>
 
<partinfo>BBA_K1976055 SequenceAndFeatures</partinfo>

Revision as of 02:56, 28 October 2016

trypsin fragment of Colicin E2 with B0034 and T7 promoter

This part encodes the trypsin 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.

Characteristics

The DNase trypsin fragment of colicin E2 consists of 137 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. This fragment differs to the part 50 in 5 additional amino acids since a trypsin digestion described in the literature leaded to this fragment. So we wanted to check out any difference in the activities between the DNase fragment with and without theese additional amino acids. 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 84
  • 1000
    COMPATIBLE WITH RFC[1000]


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