Coding

Part:BBa_K1365003:Design

Designed by: Sandra Mous   Group: iGEM14_Groningen   (2014-09-24)
Revision as of 02:23, 18 October 2014 by S.Mous (Talk | contribs) (→‎Design Notes)


NisC


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Used primers:

forward - 5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGATGAATAAAAAAAATATAAA-3'

reverse - 5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATCATTTCCTCTTCCCTC-3'

to PCR NisC from the genome and add the BioBrick prefix and suffix.


During the making of this BioBrick a mutation occured in the gene. The mutation occured at residue 185, where an AAT was changed to an AAA, changing an asparagine to a lysine. The changed amino acid was on the outside of the protein, so may not have an effect on the function of NisC.

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