PART COLLECTION

One of our approaches to improving the binding affinity between BlcR and DNA was to create new variants of the blc operator sequence. As a result, our part collection consists of 23 DNA oligos capable of binding our protein of interest BlcR to varying degrees. Together with the general experimental design and results of module 3, as well as the overview given on our libraries page, we believe these Parts may act as a framework for future iGEM teams that focus on transcription factors. For example, the different ways in which we manipulated the wildtype oligo may serve as a first step in other teams' design phase of transcription factor-binding DNA sequences, and our electrophoresis experiments can be adapted to show the functionality of a multitude of DNA-binding proteins.

Additionally, the last two oligos are the result of our partnership with the DTU iGEM 2022 team. These parts were created through an alternative approach compared to the others; instead of adjusting the wildtype sequence, the family of the blc operator was analyzed for variants. From this analysis, the overall consensus sequence of the family (oligo 21) and the operator sequence from a different A. tumefaciens strain (oligo 22) were added to our Parts. Specifically, we have chosen the consensus sequence as our favorite Basic Part (see our Parts page). The results of module 3 showed that BlcR binds significantly more strongly to this sequence than to the wildtype. Moreover, the analysis of the blc operator family showed an unexpected pattern of conservation, which may suggest a yet unknown binding mechanism of BlcR to DNA.

For further information on the biology, design, and experimental results of each Part, please be referred to the individual BioBrick pages on the Parts Registry by clicking the Part name in the table below.