Difference between revisions of "Part:BBa K1365006:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | Used primers: | + | Used primers:<br> |
| − | + | forward - 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTATAAAATTTTAATAGT-3'<br> | |
| − | forward - 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTATAAAATTTTAATAGT-3' | + | reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACTTTTTTATTTTTA-3'<br> |
| − | + | ||
| − | reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACTTTTTTATTTTTA-3' | + | |
| − | + | ||
to PCR NisR and NisK from the genome and add the BioBrick prefix and suffix. | to PCR NisR and NisK from the genome and add the BioBrick prefix and suffix. | ||
Latest revision as of 02:41, 18 October 2014
NisR and NisK
- 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
Used primers:
forward - 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTATAAAATTTTAATAGT-3'
reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTACTTTTTTATTTTTA-3'
to PCR NisR and NisK from the genome and add the BioBrick prefix and suffix.
The GTG start codon of NisR was changed into a ATG, as this is standard with most BioBricks.
During the making of this BioBrick two mutations occured in the gene. At residue 199 of NisR a GAA was changed to a GAG, not changing the amino acid at this position. At residue 300 of NisK an ACA was changed to an ATA, changing a threonine to an isoleucine.
Source
Obtained by PCR on the genome of Lactococcus lactis NZ9700, a nisin producing strain.
References
Trmčić, A. et al. (2011) Complete nisin A gene cluster from Lactococcus lactis M78 (HM219853) – obtaining the nucleic acid sequence and comparing it to other published nisin sequences. Genes Genom. 33: 217-221