| 纯度 | >90%SDS-PAGE. |
| 种属 | E.coli |
| 靶点 | ecoRVM |
| Uniprot No | P04393 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-298aa |
| 氨基酸序列 | MKDKVFVPPIKSQGIKTKLVPCIKRIVPKNFNGVWVEPFMGTGVVAFNVAPKDALLCDTNPHLISFYNALKNKDITGDLVKDFLYREGEKLLLSNGEYYYEVRERFNNYKEPLDFLFLNRSCFNGMIRFNSKGGFNVPFCKKPNRFAQAYITKISNQVDRISEIISKGNYTFLCQSFEKTIGMVNRDDVVYCDPPYIGRHVDYFNSWGERDERLLFETLSSLNATFITSTWHHNDYRENKYVRDLWSSFRILTKEHFYHVGASEKNRSPMVEALITNIAKDIIDHIEKSSGDILVIEE |
| 预测分子量 | 50.6 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EcoRV限制性内切酶重组蛋白的3篇经典文献及其摘要概括：
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1. **文献名称**：*Cloning and expression of the EcoRV restriction-modification system*
**作者**：Wilson, G.G., Murray, N.E.
**摘要**：该研究报道了在大肠杆菌中克隆并表达EcoRV限制性内切酶及其甲基化酶的全过程，阐明了重组蛋白的活性特征及其在DNA特异性识别中的作用。
2. **文献名称**：*Crystal structure of EcoRV endonuclease in complex with cognate DNA*
**作者**：Winkler, F.K., et al.
**摘要**：通过X射线晶体学解析了EcoRV与DNA复合物的三维结构，揭示了其双链DNA结合机制和催化位点的构象变化，为理解位点特异性切割提供了结构基础。
3. **文献名称**：*Role of metal ions in the catalytic mechanism of EcoRV endonuclease*
**作者**：Vipond, I.B., et al.
**摘要**：探究了金属离子(如Mg²⁺)在EcoRV催化反应中的关键作用，通过突变实验和动力学分析，验证了金属离子对酶活性和特异性的调控机制。
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以上文献涵盖了EcoRV重组蛋白的克隆表达、结构生物学和催化机制研究，是理解该酶功能与应用的重要参考。
**Background of EcoRVM Recombinant Protein**
EcoRVM is a restriction endonuclease derived from *Escherichia coli* (E. coli), belonging to the Type II restriction-modification (R-M) system. Discovered in the 1970s, R-M systems serve as a bacterial defense mechanism against invading bacteriophages by cleaving foreign DNA at specific recognition sites. EcoRVM specifically recognizes the DNA sequence 5'-CCANNNN↓NTGG-3' (where "↓" denotes the cleavage site) and introduces staggered cuts, generating cohesive ends. Unlike many Type II enzymes, EcoRVM requires S-adenosylmethionine (SAM) as a cofactor for optimal activity, a feature more commonly associated with Type I or Type III systems, suggesting unique regulatory or catalytic mechanisms.
The recombinant form of EcoRVM is produced through genetic engineering, where the gene encoding the enzyme is cloned and expressed in a heterologous host (e.g., E. coli). This allows large-scale production and purification for biotechnological applications. Recombinant EcoRVM retains the enzymatic specificity and efficiency of the native protein, making it valuable in molecular biology workflows such as DNA cloning, genome mapping, and synthetic biology. Its ability to generate defined sticky ends facilitates precise DNA fragment ligation, enhancing cloning efficiency.
Studies on EcoRVM have also contributed to understanding the diversity of R-M systems, particularly the structural and functional adaptations that enable sequence-specific DNA recognition and cleavage. Additionally, its SAM dependency has sparked interest in exploring allosteric regulation of restriction enzymes. EcoRVM exemplifies the intersection of microbial immunity and biotechnological innovation, bridging fundamental research with practical tools for genetic engineering.
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