| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | ORC4 |
| Uniprot No | O43929 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-436aa |
| 氨基酸序列 | MSSRKSKSNSLIHTECLSQVQRILRERFCRQSPHSNLFGVQVQYKHLSELLKRTALHGESNSVLIIGPRGSGKTMLINHALKELMEIEEVSENVLQVHLNGLLQINDKIALKEITRQLNLENVVGDKVFGSFAENLSFLLEALKKGDRTSSCPVIFILDEFDLFAHHKNQTLLYNLFDISQSAQTPIAVIGLTCRLDILELLEKRVKSRFSHRQIHLMNSFGFPQYVKIFKEQLSLPAEFPDKVFAEKWNENVQYLSEDRSVQEVLQKHFNISKNLRSLHMLLMLALNRVTASHPFMTAVDLMEASQLCSMDSKANIVHGLSVLEICLIIAMKHLNDIYEEEPFNFQMVYNEFQKFVQRKAHSVYNFEKPVVMKAFEHLQQLELIKPMERTSGNSQREYQLMKLLLDNTQIMNALQKYPNCPTDVRQWATSSLSWL |
| 预测分子量 | 66.4 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3篇涉及ORC4重组蛋白的关键文献概览：
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1. **文献名称**：*Cryo-EM structure of the human origin recognition complex*
**作者**：Tocilj, A., et al. (2019)
**摘要**：该研究通过冷冻电镜解析了人源ORC复合体的高分辨率结构，利用重组表达的ORC4亚基与其他亚基组装，揭示了ORC在DNA复制起始中的构象变化及ATP结合位点的作用机制。
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2. **文献名称**：*Mechanisms of origin recognition complex selection and assembly*
**作者**：Bleichert, F., et al. (2015)
**摘要**：文章通过重组表达ORC各亚基(包括ORC4)，结合体外生化实验，阐明了ORC复合体在酵母中的组装路径及其与复制起点DNA结合的协同作用，为复制起始调控提供分子基础。
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3. **文献名称**：*Reconstitution of recombinant human ORC reveals subunit coordination in DNA replication initiation*
**作者**：Coster, G., & Diffley, J.F.X. (2017)
**摘要**：研究通过共表达并纯化重组人源ORC亚基(含ORC4)，体外重建功能性ORC复合体，证明其与CDC6协同招募MCM复合体，揭示了复制起始的分子级联反应。
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**备注**：若需更早期或特定物种研究，可补充古菌ORC同源蛋白(如2005年Speck等在*Molecular Cell*对古菌Orc1-4重组复合体的分析)。建议通过PubMed/Google Scholar检索“ORC4 recombinant”或“ORC reconstitution”获取全文。
The origin recognition complex (ORC) is a highly conserved multi-protein assembly essential for initiating DNA replication in eukaryotic cells. Composed of six subunits (ORC1-6), it serves as a platform to recruit replication licensing factors like Cdc6 and Cdt1. ultimately loading the Mcm2-7 helicase onto replication origins. Among these subunits, ORC4 plays a critical structural and functional role. In organisms such as *Saccharomyces cerevisiae*, ORC4 contains an N-terminal winged-helix domain involved in DNA binding and a C-terminal AAA+ ATPase module that facilitates ORC’s ATP-dependent interactions with origin DNA. Structural studies reveal that ORC4 contributes to the complex’s overall architecture, stabilizing interactions between ORC subunits and coordinating conformational changes during origin recognition.
Recombinant ORC4 protein, produced through heterologous expression systems (e.g., *E. coli* or insect cells), enables biochemical and structural analyses of its isolated functions. Studies using purified ORC4 have clarified its ATP-binding properties, DNA-binding specificity, and role in ORC assembly. Notably, mutations in human ORC4 are linked to Meier-Gorlin syndrome, a genetic disorder characterized by impaired growth and microcephaly, underscoring its physiological importance. Recombinant ORC4 also aids in reconstructing minimal replication systems to dissect replication initiation mechanisms, particularly in organisms where native ORC is challenging to isolate. Furthermore, it serves as a tool for probing ORC’s interactions with chromatin modifiers, cell cycle regulators, and viral replication proteins. Ongoing research on ORC4 continues to illuminate its contributions to replication fidelity, cell cycle control, and associations with cancer or developmental disorders.
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