Team:KAIT Japan/Basic Part

Parts


We designed four iGEM parts in the year.



    • BBa_K1740000
    This part is the open reading frame of optimized Dronpa 145N to E.coli. Dronpa 145N mutated wild-Dronpa which changed from 145Lys to Asn. Dronpa 145N switched on under 390nm light from a non-fluorescent state into a fluorescent state, and back again under 503nm light. At the fluorescent state, Dronpa 145N forms a tetramer which is combined with other Dronpa 145N. On the other hand, at the non-fluorescent state, Dronpa 145N forms a monomer. This is reversible action, and it is controlled by light. Dronpa 145N is one of the Green Fluorescent Proteins, it is in its on-state, displaying bright green fluorescence.



    • BBa_K1740001
    This part is Dronpa 145K ORF. Dronpa 145K is similar to wild-type Dronpa. Dronpa 145K is Dronpa 145N which changed from 145Asn to 145Lys. And it has optimized to E.coli. Dronpa 145N is displaying bright green fluorescence by forming a tetramer, but Dronpa 145K brights green by a monomer. However, Dronpa 145K switched on under 390nm light from a non-fluorescent state into a fluorescent state, and back again under 503nm light too.



    • BBa_K1740003
    This part contains pLac + RBS + Dronpa 145N +Double stop codon. The coding sequence Dronpa145N contained in the part: BBa_K1740000 doesn't contain the stop codon to the terminus. In that part, it doesn't contain a promoter sequence and a ribosomal binding site(RBS) sequence necessary for the protein expression. Therefore, we designed this part which added to Lac promoter(pLac), RBS and double stop codon to Dronpa 145N.



    • BBa_K1740004
    This part contains the coding sequence for Dronpa 145N and Multi Cloning Sites(MCS). This is designed so that Dronpa 145N is tagged to both terminus of the target protein. MCS contains the cleavage site of next restriction enzymes: Xho I, Hind III, Nde I, Kpn I, Sal I, Sac I and BamH I. We can obtain a fusion protein to react with A target DNA fragment which reacted their restriction enzymes and this part that treated restriction enzyme same as the fragment. The target protein activity in the fusion protein can control by light.



    BBa_K1740000, 001, 003 were submitted. But, BBa_K1740004 wasn’t submitted.