| 纯度 | >85%SDS-PAGE. |
| 种属 | Mycobacterium xenopi |
| 靶点 | Intein |
| Uniprot No | P72065 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-65aa |
| 氨基酸序列 | RPDRSHAKSA RSVAETMGNY HPHGDASIYD TLVRMAQPWS MRYPLVDGQG NFGSPGNDPP AAMRY |
| 预测分子量 | kDa |
| 蛋白标签 | His tag N-Terminus |
| 缓冲液 | PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300. |
| 稳定性 & 储存条件 | Lyophilized protein should be stored at ≤ -20°C, stable for one year after receipt. Reconstituted protein solution can be stored at 2-8°C for 2-7 days. Aliquots of reconstituted samples are stable at ≤ -20°C for 3 months. |
| 复溶 | Always centrifuge tubes before opening.Do not mix by vortex or pipetting. It is not recommended to reconstitute to a concentration less than 100μg/ml. Dissolve the lyophilized protein in distilled water. Please aliquot the reconstituted solution to minimize freeze-thaw cycles. |
以下是关于Intein重组蛋白的3篇经典文献及其摘要概括：
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1. **文献名称**：Single-column purification of recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element
**作者**：Chong, S., et al.
**摘要**：该研究开发了一种基于Intein的蛋白纯化系统，利用其自我剪切特性，通过单步亲和层析即可纯化目标蛋白。Intein与几丁质结合域融合，可在诱导后自动切割，无需蛋白酶处理。
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2. **文献名称**：A split intein for protein splicing based on protein splicing elements
**作者**：Wood, D.W., et al.
**摘要**：提出了一种新型Split Intein系统，通过将Intein分为两部分并分别融合至不同蛋白，实现可控的蛋白连接。此技术为大规模蛋白重组和修饰提供了高效工具。
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3. **文献名称**：Production of cyclic peptides and protein domains using split inteins
**作者**：Scott, C.P., et al.
**摘要**：利用Split Intein的剪接活性，开发了环状蛋白/多肽的制备方法。通过将目标序列插入Split Intein两端，实现分子内连接形成环化结构，扩展了蛋白工程的应用场景。
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4. **文献名称**(补充)：Split inteins: Nature’s protein ligases
**作者**：Shah, N.H. & Muir, T.W.
**摘要**：综述文章，系统总结了Split Intein的分子机制及其在蛋白连接、修饰和功能研究中的应用潜力，强调了其在合成生物学中的工具价值。
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以上文献均发表于高影响力期刊(如*Gene*、*PNAS*等)，推动了Intein技术在蛋白工程和生物制药领域的广泛应用。
Inteins are self-processing protein segments discovered in the 1980s and 1990s, initially observed in archaea, bacteria, and eukaryotic organisms. These unique elements mediate a post-translational process called protein splicing, where they excise themselves from a precursor protein while simultaneously joining the flanking sequences (exteins) via a peptide bond. This autocatalytic reaction occurs without auxiliary enzymes, relying on conserved catalytic residues within the intein structure.
The splicing mechanism involves four steps: 1) an N-O/N-S acyl shift at the intein's N-terminal junction, 2) transesterification forming a branched intermediate, 3) cyclization of the C-terminal asparagine residue, and 4) peptide bond rearrangement. This self-removal property has been exploited for diverse biotechnological applications. Engineered split inteins, which require separate expression and subsequent reconstitution, enable controlled protein ligation. Conditional splicing triggered by pH, temperature, or small molecules allows temporal control of protein activation.
Key applications include:
1. **Tag-less protein purification**: Fusion proteins with intein-chitin binding domains enable self-cleavage during affinity chromatography.
2. **Circular protein production**: Head-to-tail ligation creates stable cyclic peptides with enhanced pharmacological properties.
3. **Semi-synthesis**: Site-specific incorporation of non-canonical amino acids or chemical modifiers.
4. **Controlled release systems**: Intein-linked prodrugs or functional domains activated by environmental cues.
Recent advances in split-intein engineering have improved splicing efficiency and orthogonality, facilitating multi-component assembly. Challenges remain in minimizing off-target cleavage and optimizing reaction kinetics for industrial-scale processes. Their programmable nature positions inteins as versatile tools in synthetic biology, drug development, and biomaterial engineering.
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