VectorBuilderを使って自分のベクターをデザインすることができます。以下によく使われるコンポーネントの解説があります。
注:
複数のORFを1つのプロモーターで制御するために使われるリンカーには2種類あります:
複数のORFを1つのプロモーターで制御するために使われるリンカーには2種類あります:
- read-throughリンカー （例 融合リンカーや2Aペプチドなど）:上流のORFにストップコドンがない場合、翻訳はリンカーを挟んで上流ORFから下流ORFまで一続きに起こります。
- 非read-throughリンカー（例 IRES）:上流のORFがストップコドンで終了している場合、リンカーの直前で翻訳が停止します。
Fusion (リードスルー)
| 名前 | 詳細 | アプリケーションノート | レファレンス | シークエンス |
|---|---|---|---|---|
| 3xGGGGS | 3 tandem GGGGS | Commonly used flexible linker for fusing two proteins. | Adv Drug Deliv Rev. 65: 1357 (2013) | View |
| 3xGS | 3 tandem GS | Flexible linker for fusing two proteins. | Protein Sci. 22:153 (2013) | View |
2Aペプチド (リードスルー)
| 名前 | 詳細 | アプリケーションノート | レファレンス | シークエンス |
|---|---|---|---|---|
| T2A | Self-cleaving 2A peptide from Thosea asigna virus | Causes co-translational cleavage of the encoded polypeptide. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by 2A. P2A and T2A have higher cleavage efficiency compared to other 2As. | PLoS One. 6:e18556 (2011) | View |
| P2A | Self-cleaving 2A peptide from Porcine teschovirus-1 | Causes co-translational cleavage of the encoded polypeptide. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by 2A. P2A and T2A have higher cleavage efficiency compared to other 2As. | PLoS One. 6:e18556 (2011) | View |
| F2A | Self-cleaving 2A peptide from foot-and-mouth disease virus | Causes co-translational cleavage of the encoded polypeptide. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by 2A. | PLoS One. 6:e18556 (2011) | View |
| E2A | Self-cleaving 2A peptide from Equine rhinitis A virus | Causes co-translational cleavage of the encoded polypeptide. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by 2A. | PLoS One. 6:e18556 (2011) | View |
| T2A(dme) | Self-cleaving 2A peptide from Thosea asigna virus, codon-optimized for Drosophila melanogaster. | Causes co-translational cleavage of the encoded polypeptide. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by 2A. P2A and T2A have higher cleavage efficiency compared to other 2As. | PLoS One. 9:e100637 (2014); Genetics. 190:1139 (2012) | View |
| P2A(dme) | Self-cleaving 2A peptide from Porcine teschovirus-1, codon-optimized for Drosophila melanogaster. | Causes co-translational cleavage of the encoded polypeptide. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by 2A. P2A and T2A have higher cleavage efficiency compared to other 2As. | PLoS One. 9:e100637 (2014); Genetics. 190:1139 (2012) | View |
| E2A(dme) | Self-cleaving 2A peptide from Equine rhinitis A virus, codon-optimized for Drosophila melanogaster. | Causes co-translational cleavage of the encoded polypeptide. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by 2A. | PLoS One. 9:e100637 (2014); Genetics. 190:1139 (2012) | View |
Internal Ribosome Entry Sites (非リードスルー)
| 名前 | 詳細 | アプリケーションノート | レファレンス | シークエンス |
|---|---|---|---|---|
| IRES | Encephalomyocarditis virus internal ribosome entry site | Recruits ribosome to initiate translation internally on a transcript independent of its 5' end. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by IRES. | Biotechniques. 41:283 (2006) | View |
| Ubx-IRES | Drosophila melanogaster Ultrabithorax internal ribosome entry site | Recruits ribosomes to initiate translation internally on a transcript independent of its 5' end. Multiple proteins can be made from a polycistronic transcript containing multiple ORFs separated by IRES. | Development. 122:747 (1996); The EMBO Journal. 6:1775 (1987) | View |