Difference between revisions of "Part:BBa K1659210:Design"
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This part is a modified version of [https://parts.igem.org/Part:BBa_K1659200:Design BBa_K1659200], where thymine 582 was mutated into a cytosine, conserving the His-194 which it encodes for. The site-directed mutagenesis was performed using a modified [https://www.neb.com/applications/cloning-and-synthetic-biology/site-directed-mutagenesis NEB Q5 Mutagenesis Protocol], which is fully described in our team's [http://2015.igem.org/Team:Oxford/Protocols Protocols Page]. | This part is a modified version of [https://parts.igem.org/Part:BBa_K1659200:Design BBa_K1659200], where thymine 582 was mutated into a cytosine, conserving the His-194 which it encodes for. The site-directed mutagenesis was performed using a modified [https://www.neb.com/applications/cloning-and-synthetic-biology/site-directed-mutagenesis NEB Q5 Mutagenesis Protocol], which is fully described in our team's [http://2015.igem.org/Team:Oxford/Protocols Protocols Page]. | ||
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<img src="https://static.igem.org/mediawiki/parts/c/c8/Oxford15_DspBQC1.png" alt="DspB quikchange" width="345" height="275" style="float:right"> | <img src="https://static.igem.org/mediawiki/parts/c/c8/Oxford15_DspBQC1.png" alt="DspB quikchange" width="345" height="275" style="float:right"> | ||
<figcaption> | <figcaption> | ||
− | <p style="font-size: | + | <p style="font-size:13px"> |
− | The mutation created was necessary for the removal of the BspHI restriction site found within the coding sequence of BBa_K1659200. This is to help facilitate the characterization of the part, as we characterize our coding sequence parts by cloning them into the commercial expression vector | + | The mutation created was necessary for the removal of the BspHI restriction site found within the coding sequence of BBa_K1659200. This is to help facilitate the characterization of the part, as we characterize our coding sequence parts by cloning them into the commercial expression vector <a href="http://www.thermofisher.com/order/catalog/product/V43001">pBAD/HisB</a>, the process of which requires restriction digest at the BspHI site upstream of the part added on via PCR for ligation with the NcoI site found on the vector. |
<br> | <br> | ||
<br> | <br> | ||
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</figure> | </figure> | ||
</html> | </html> | ||
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Latest revision as of 10:21, 14 September 2015
Dispersin B with mutation at nucleotide 582 (T to C)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 262
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 418
Design Notes and Sources
This part is a modified version of BBa_K1659200, where thymine 582 was mutated into a cytosine, conserving the His-194 which it encodes for. The site-directed mutagenesis was performed using a modified NEB Q5 Mutagenesis Protocol, which is fully described in our team's [http://2015.igem.org/Team:Oxford/Protocols Protocols Page].
References
[1] SnapGene software (from GSL Biotech; available at snapgene.com)