BioBricks made by iGEM Stockholm 2016

On this page, we compiled an overview of all the BioBricks we have created this summer in our mission to degrade biofilms. The links below will lead you the corresponding registry entry, if you'd like to to know more about the proteins, have a look at our project description or our results page.

Composite Parts

The composite parts in our project were all assembled in a similar fashion, creating two new BioBricks for each protein. The first construct was generally a simple assembly with a T7 promoter allowing for inducible protein expression. The second BioBrick contained an additional sequence consisting or a linker and two tags (LT) - one needed for purification and one for Sortase-A-mediated conjugation of our proteins to spider silk.

BBa_K2144000 T7-Nuclease
BBa_K2144011 T7-Nuclease-LT

Nuc2, isolated form S. epidermidis is a well-known nuclease that we assembled with a T7 promoter in order to degrade biofilm-associated extracellular DNA.

BBa_K2144001 T7-Lysostaphin
BBa_K2144005 T7-Lysostaphin-LT

Lysostaphin displays a highly specific endopeptidase activity which has been shown to degrade a key components of many biofilms as well as peptidoglycans found in cell walls.

BBa_K2144010 T7-Esp
BBa_K2144012 T7-Esp-LT

Esp is a serine protease secreted by S. epidermidis which degrades a variety of proteins associated with cell walls or the biofilm itself, among them many virulence factors.

BBa_K2144003 T7-Defensin
BBa_K2144007 T7-Defensin-LT

Defensins are a class of molecules known to disrupt membrane integrity of pathogenic organisms, dramatically increasing membrane permeability and leading to cell death.

Basic Parts

The basic part we worked on was Sortase A; a transpeptidase commonly used in biotechnology for the conjugation of peptides.

BBa_K2144008 Sortase A

Part Collection - The Sortase Package

Sortase A alone is a useful tool but we aimed to provide an entire toolbox, containing the Sortase A BioBrick as well as a vector for simple addition of Sortase tags to any protein and user-friendly instructions for the assembly of proteins using this toolbox. More information about the package can be found here.
BBa_K2144008 Sortase A
BBa_K2144101 LT

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