Difference between revisions of "Part:BBa K1859020"
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We designed this generator[K1859020] to express long-chain protein with keeping functionality of the protein.
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We designed this generator[K1859020] to express long-chain protein with keeping functionality of the protein, giving the red fluorescence.
 
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The generator consists of リンカーを結合させたサーキュラーパーツ([BBa_]と[BBa_])、His-RFP without stop codon[BBa_K1332002]、Lacl[]、RBS[]、DT[].
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The generator consists of Circular parts combined the linker(
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<a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_K1859007" > [BBa_K1859007] </a>
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and
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<a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_K1859011" > [BBa_K1859011] </a>
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), His-RFP without stop codon
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<a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_BBa_K1332002" >[BBa_K1332002] </a, Lacl
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<a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_R0010" >[BBa_R0010] </a>
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, RBS
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<a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_R0034" >[BBa_B0034] </a>
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and DT
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<a href= "https://parts.igem.org/wiki/index.php?title=Part:BBa_R0010" >[BBa_B0015] </a>
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.
 
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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。
 
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。

Revision as of 05:00, 18 September 2015

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。