リンカー

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:
  • 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。
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