Part:BBa_K1859020

Histidine tag and RFP linker [GGSGGS*2] semi-permanent generator

We designed this generator[K1859020] to express long-chain protein with keeping functionality of the protein, giving the red fluorescence.

In the study of iGEM Gifu 2014, we made long-chain protein from Circular mRNA, but it lost functionality. It is thought that folding of the protein failed for a cause．

Therefore we designed some linkers to fix the folding of the protein. Then, we combined circular parts, the 3'side of the intron [BBa_K1332005] and the 5'side of the intron [BBa_K1332003] , with those linkers and made them parts. The generator consists of Circular parts combined the linker( [BBa_K1859007] and [BBa_K1859011] ), His-RFP without stop codon [BBa_K1332002] [BBa_R0010] , RBS [BBa_B0034] and DT [BBa_B0015] .

しかし、このgeneratorを導入した大腸菌で発現させたRFPは、蛍光を発するどころか、タンパク質の長鎖化さえされなかった。

原因として、私たちは導入したリンカーの配列が、スプライシングに関するリボザイムの構造を変化させ、機能が消失したためであると考えている。