Team:UCopenhagen/Improve
Improved biobrick
We have improved the function of a previous Biobrick: Original biobrick; http://parts.igem.org/Part:BBa_K864100. Our improved biobrick: http://parts.igem.org/Part:BBa_K2455002
The This is an improved version of the Super Yellow Fluorescent Protein 2 (BBa_K864100) carrying an N-terminal nona-arginine tag,
and a C-terminal hexa-histidine tag. The R9-tag is a Cell-Penetrating Peptide giving it's associated protein the ability to pass through plasma membranes.
We made the improvement by inserting BBa_K864100 into the USER casette of our third biobrick, the Cell-Penetrating USER Cassette http://parts.igem.org/Part:BBa_K2455003.
This biobrick contains an AsiSI/Nb.BSMI USER Cassette with a N-terminal nona-arginine (R9) Cell-penetrating peptide.
The nona-arginine (R9) gives any cassette inserted proteins the ability to pass through plasma membranes.
In addition, the USER Cassette carries a N-terminal hexa-histidine tag which allows for easy purification.
For our study we demonstrated that proteins inserted into the cassette can be purified using the histidine-tag,
and subsequently used to stain E. coli and Pseudomonas putida cells, presumably through internalization caused by the R9 peptide.
