Team:Groningen/Template/MODULE/Notebook/toolbox/week1

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