| 纯度 | >90%SDS-PAGE. |
| 种属 | E.coli |
| 靶点 | NSP12 |
| Uniprot No | Q08534 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-932aa |
| 氨基酸序列 | SADAQSFLNRVCGVSAARLTPCGTGTSTDVVYRAFDIYNDKVAGFAKFLKTNCCRFQEKDEDDNLIDSYFVVKRHTFSNYQHEETIYNLLKDCPAVAKHDFFKFRIDGDMVPHISRQRLTKYTMADLVYALRHFDEGNCDTLKEILVTYNCCDDDYFNKKDWYDFVENPDILRVYANLGERVRQALLKTVQFCDAMRNAGIVGVLTLDNQDLNGNWYDFGDFIQTTPGSGVPVVDSYYSLLMPILTLTRALTAESHVDTDLTKPYIKWDLLKYDFTEERLKLFDRYFKYWDQTYHPNCVNCLDDRCILHCANFNVLFSTVFLPTSFGPLVRKIFVDGVPFVVSTGYHFRELGVVHNQDVNLHSSRLSFKELLVYAADPAMHAASGNLLLDKRTTCFSVAALTNNVAFQTVKPGNFNKDFYDFAVSKGFFKEGSSVELKHFFFAQDGNAAISDYDYYRYNLPTMCDIRQLLFVVEVVDKYFDCYDGGCINANQVIVNNLDKSAGFPFNKWGKARLYYDSMSYEDQDALFAYTKRNVIPTITQMNLKYAISAKNRARTVAGVSICSTMTNRQFHQKLLKSIAATRGATVVIGTSKFYGGWHNMLKTVYSDVENPHLMGWDYPKCDRAMPNMLRIMASLVLARKHTTCCSLSHRFYRLANECAQVLSEMVMCGGSLYVKPGGTSSGDATTAYANSVFNICQAVTANVNALLSTDGNKIADKYVRNLQHRLYECLYRNRDVDTDFVNEFYAYLRKHFSMMILSDDAVVCFNSTYASQGLVASIKNFKSVLYYQNNVFMSEAKCWTETDLTKGPHEFCSQHTMLVKQGDDYVYLPYPDPSRILGAGCFVDDIVKTDGTLMIERFVSLAIDAYPLTKHPNQEYADVFHLYLQYIRKLHDELTGHMLDMYSVMLTNDNTSRYWEPEFYEAMYTPHTVLQ |
| 预测分子量 | 108.2 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SARS-CoV-2 NSP12重组蛋白的3篇代表性文献(内容基于真实研究概括，但文献标题和作者为示例性信息)：
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1. **文献名称**: *Structural basis for RNA replication by the SARS-CoV-2 polymerase*
**作者**: Yin, W. et al.
**摘要**: 本研究解析了重组表达的NSP12(RdRp)与辅助因子NSP7/NSP8复合体的冷冻电镜结构，揭示了其RNA依赖的RNA聚合酶活性机制，并证明其与瑞德西韦等抗病毒药物的相互作用。
2. **文献名称**: *Characterization of recombinant SARS-CoV-2 RNA-dependent RNA polymerase activity*
**作者**: Subissi, L. et al.
**摘要**: 报道了在大肠杆菌中重组表达并纯化NSP12蛋白的方法，验证其与NSP7/NSP8形成的复合体在体外的RNA聚合酶活性，为抗病毒药物筛选提供实验模型。
3. **文献名称**: *Cryo-EM structure of the SARS-CoV-2 RNA replication complex*
**作者**: Hillen, H.S. et al.
**摘要**: 通过重组表达系统获得NSP12-NSP7-NSP8复合体，结合冷冻电镜技术解析其高分辨率结构，阐明病毒RNA复制过程中模板识别与核苷酸催化的分子机制。
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**备注**：以上文献标题和内容基于2020-2022年相关领域研究的典型方向，实际文献需通过PubMed或Google Scholar以关键词“SARS-CoV-2 NSP12 recombinant”查询具体信息。
**Background of NSP12 Recombinant Protein**
Nonstructural protein 12 (NSP12), the RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2. is a central enzyme in viral replication and transcription. As a key component of the replication-transcription complex (RTC), NSP12 catalyzes the synthesis of viral RNA using the genomic RNA template, enabling the virus to replicate its genome within host cells. Its critical role in the viral life cycle makes it a prime target for antiviral drug development, notably exemplified by remdesivir, a nucleotide analog that inhibits RdRp activity.
NSP12 functions in conjunction with accessory factors NSP7 and NSP8. which enhance its processivity and stability. Structural studies using cryo-electron microscopy have revealed that NSP12 adopts a conserved RdRp architecture, resembling a right hand with fingers, palm, and thumb domains. Unique to coronaviruses, NSP12 possesses an N-terminal extension that interacts with viral RNA and contributes to its enzymatic activity. The recombinant form of NSP12 is typically produced in heterologous expression systems (e.g., *E. coli* or insect cells*) for biochemical and structural studies. Purification often involves affinity chromatography and tag removal to obtain functional, soluble protein.
Research on recombinant NSP12 has advanced understanding of its mechanism, including primer-template RNA binding, nucleotide incorporation, and proofreading via exonuclease activity (mediated by NSP14). These insights are vital for designing broad-spectrum antivirals targeting conserved RdRp regions. Additionally, recombinant NSP12 serves as a tool for high-throughput drug screening and investigating resistance mutations. Its study not only illuminates coronavirus replication but also informs strategies to combat emerging RNA viruses.
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