Difference between revisions of "Part:BBa K1413043"
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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. | 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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===Usage and Biology=== | ===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. | ||
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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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===References=== | ===References=== | ||
-Bender, J., & Kleckner, N. (1988). Genetic Evidence That TnlO Transposes by a Nonreplicative Mechanism, 45, 801–815 | -Bender, J., & Kleckner, N. (1988). Genetic Evidence That TnlO Transposes by a Nonreplicative Mechanism, 45, 801–815 | ||
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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). | -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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 244
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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).