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元件指南
VectorBuilder提供了許多常用的載體元件，可供用戶在設(shè)計載體時進(jìn)行選擇。下表按類型提供了這些常用元件的詳細(xì)信息。
備注:
連接子（linker）用于連接由單個啟動子驅(qū)動的多個ORF同時表達(dá)。連接子（linker）有兩種類型：
連接子（linker）用于連接由單個啟動子驅(qū)動的多個ORF同時表達(dá)。連接子（linker）有兩種類型：
- 通讀型連接子（例如融合連接子或2A肽）：上游ORF缺少終止密碼子，使得翻譯從上游ORF通過連接子進(jìn)入下游的ORF。
- 非通讀型連接子（例如IRES）：上游ORF以終止密碼子結(jié)束，使得上游ORF的翻譯在到達(dá)接頭之前終止。
融合蛋白（通讀型連接子）
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| 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肽（通讀型連接子）
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| 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 |
IRES（非連續(xù)翻譯）
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| 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 |
