Difference between revisions of "Part:BBa K1413043"

 
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<partinfo>BBa_K1413043 short</partinfo>
 
<partinfo>BBa_K1413043 short</partinfo>
  
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This part is the coding sequence for the transposase Tn10.
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This part is the coding sequence for the transposase Tn10. A cut-and-paste mechanism where the transposase allows insertion of a transposon randomly in the genome. The transposon must be surrounded by IS10 sequences.
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This transposase allows insertion of transposon randomly in the genome. The transposon must be surrounded by IS10 sequences.
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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This part was extracted from pNK2 plasmid (see BBa_K1413044). This part was used to insert the transposon in the genome of bacteria like E. coli and Pseudovibrio denitrificans.
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<html>
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<img src="https://static.igem.org/mediawiki/parts/c/ca/Kanamycine_gel_%281%29.png" width=50%/>
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</html>
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Picture of an electrophoresis gel of the PCR which amplifies the kanamycine resistance cassette. The template of this PCR was Pseudovibrion denetrificans genome after electroporation of the transposon plasmid pNK2, control was P. denetrificans without electroporation. As expected, all clones have the insertion of the transposon, except the control.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1413043 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1413043 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1413043 parameters</partinfo>
 
<partinfo>BBa_K1413043 parameters</partinfo>
 
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===References===
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-Bender, J., & Kleckner, N. (1988). Genetic Evidence That TnlO Transposes by a Nonreplicative Mechanism, 45, 801–815
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<br>
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-Crellin, P., & Chalmers, R. (2001). Protein±DNA contacts and conformational changes in the Tn 10 transpososome during assembly and activation for cleavage, 20(14).

Latest revision as of 18:18, 2 November 2014

Transposase Tn10

This part is the coding sequence for the transposase Tn10. A cut-and-paste mechanism where the transposase allows insertion of a transposon randomly in the genome. The transposon must be surrounded by IS10 sequences.


Usage and Biology

This part was extracted from pNK2 plasmid (see BBa_K1413044). This part was used to insert the transposon in the genome of bacteria like E. coli and Pseudovibrio denitrificans.


Picture of an electrophoresis gel of the PCR which amplifies the kanamycine resistance cassette. The template of this PCR was Pseudovibrion denetrificans genome after electroporation of the transposon plasmid pNK2, control was P. denetrificans without electroporation. As expected, all clones have the insertion of the transposon, except the control.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 244
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

-Bender, J., & Kleckner, N. (1988). Genetic Evidence That TnlO Transposes by a Nonreplicative Mechanism, 45, 801–815
-Crellin, P., & Chalmers, R. (2001). Protein±DNA contacts and conformational changes in the Tn 10 transpososome during assembly and activation for cleavage, 20(14).