Team:Groningen/Template/MODULE/Notebook/toolbox/week1
From 2014.igem.org
7 July - 13 July
Primers were designed for making BioBricks out of the separate genes from
the nisin operon. For this, the sequence of the transposon
Tn5307 was used, a transposon that contains the nisin operon, and of
which the sequence is known.1 This sequence is close to the sequence
of the transposon Tn5276, the transposon which will form the template
for making the new BioBricks. The primers for making a BioBrick out of
PNisI with a RBS were based on a sequence found in a research paper that
documented the identification of this promoter.2 The list of
primersequences can be found in the first table at the bottom of this page.
It was decided to combine the genes NisR
and NisK as one BioBrick and NisF, NisE and NisG as one BioBrick, because
these genes are related in function or work together.
There were also primers designed for making a BioBrick out of the sfGFP(Bs), the primersequences can be found at second table at the bottom of this page. This sfGFP
was originally optimized for Bacillus subtilis. When it was
tested in Lactococcus lactis, the sfGFP(Bs) was shown to perform
really well in L. lactis as well.3