Difference between revisions of "Part:BBa K1659300:Design"
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| − | BLAST search using translated sequence of BBa_K729004, from which only the nucleotide sequence that coded for the DNase was identified | + | BLAST search using translated sequence of BBa_K729004, from which only the nucleotide sequence that coded for the DNase was identified and isolated |
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| − | 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 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. | + | 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. |
A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography. | A Hisx6 tag is added at the C-terminus for ease of protein purification using metal-affinity chromatography. | ||
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
BLAST search using translated sequence of BBa_K729004, from which only the nucleotide sequence that coded for the DNase was identified and isolated
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.