Difference between revisions of "Team:Kyoto/Parts"

Our best basic part is BBa_K3815000. This is composed of ELK16, PT linker, Mxe Gyr intein. It can be expressed by attaching the sequence of the targeted protein to the N-terminus of Intein. This part is used for the purification method called ELK16system, which past iGEM teams had not been used. In this ELK16system, the ELK16 part is self-condensing, so the fusion protein can be extracted only by centrifugation, and the intein part is self-cleaved by DTT, so the target protein can be easily separated. The ELK16system will help improve the technology of recombinant protein purification, although it has some problems such as the presence of many other impurities in the protein recovery process.

These parts are the improvement from BBa_K1051206, the wildtype ssrA tag. Three C-terminal amino acids LAA in WT tag are replaced in these mutants, therefore resulting in a reduced protein degradation efficiency to various degrees (see Result). The collection of these mutants consist of a large repertoire of protein degradation tags of various levels, which can be applied for fine-tuning of many kinds of synthetic systems that require a precise control of proteins.