Difference between revisions of "Part:BBa K1659300:Design"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1659300 short</partinfo> | <partinfo>BBa_K1659300 short</partinfo> | ||
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− | ===Design Notes=== | + | ===Design Notes and Sources=== |
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+ | <html> | ||
+ | <figure> | ||
+ | <img src="https://static.igem.org/mediawiki/parts/6/61/Oxford15_blastdnase.png" alt="DNase BLAST" width="856" height="123"> | ||
+ | <figcaption> | ||
+ | <p style="font-size:11px"> | ||
+ | BLAST search using translated sequence of BBa_K729004, from which only the nucleotide sequence that coded for the DNase was identified and isolated | ||
+ | </p> | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | </html> | ||
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− | to | + | The gene sequence for micrococcal DNase was obtained from [https://parts.igem.org/Part:BBa_K729004 BBa_K729004], a part comprising DsbA 2-19 fused to the N-terminal of micrococcal nuclease made by [http://2012.igem.org/Team:University_College_London Team UCL 2012]. The part sequence was not annotated, and as such we ran a [http://blast.ncbi.nlm.nih.gov/blast/Blast.cgi?PROGRAM=blastp&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome Protein BLAST search] using the part's translated sequence to identify the bases that coded for our desired peptide [1]. The sequence for micrococcal DNase was adopted as-is from said part, without further codon optimization. |
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+ | A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography. | ||
+ | |||
+ | |||
===References=== | ===References=== | ||
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+ | [1] Stephen F. Altschul, Thomas L. Madden, Alejandro A. Schäffer, Jinghui Zhang, Zheng Zhang, Webb Miller, and David J. Lipman, 1997. "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 25:3389-3402. |
Latest revision as of 01:27, 15 September 2015
Micrococcal Nuclease (DNase)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes and Sources
The gene sequence for micrococcal DNase was obtained from BBa_K729004, a part comprising DsbA 2-19 fused to the N-terminal of micrococcal nuclease made by [http://2012.igem.org/Team:University_College_London Team UCL 2012]. The part sequence was not annotated, and as such we ran a [http://blast.ncbi.nlm.nih.gov/blast/Blast.cgi?PROGRAM=blastp&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome Protein BLAST search] using the part's translated sequence to identify the bases that coded for our desired peptide [1]. The sequence for micrococcal DNase was adopted as-is from said part, without further codon optimization.
A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography.
References
[1] Stephen F. Altschul, Thomas L. Madden, Alejandro A. Schäffer, Jinghui Zhang, Zheng Zhang, Webb Miller, and David J. Lipman, 1997. "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 25:3389-3402.