纯度 | >90%SDS-PAGE. |
种属 | Colwellia |
靶点 | dinB |
Uniprot No | Q487H6 |
内毒素 | < 0.01EU/μg |
表达宿主 | E.coli |
表达区间 | 1-352aa |
氨基酸序列 | MGNQKKIIHIDMDCFYAAIEMRDFPEYQNIPLAVGGDGPRSVLCTSNYQARQFGVRSAMPAIKAKQLCPHLKIVHGRMDVYKETSKNIREIFSRYTDLIEPLSLDEAYLDVTDATMCQGSATLIAERIRADIFNELNLTASAGIAPNKFLAKIASDENKPNGQCVITPDKVANFVEQLSLKKIPGIGPKTFEKLNRHGYVTCADVRQSNIRALQNIVGKFANSLYLKSHGVDNRDLEVSRQRKSLAIETTLAHDISTQDECKLVIDSLYQKLLTRLAPHSNREIIRQGVKLKFTDFNQTTVETQSNECQQALFISLLSKAYSRSNKRGVRLVGLTLGFADSPGESQQLSLSL |
预测分子量 | 43.3 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dinB重组蛋白的3篇参考文献,包含文献名称、作者及摘要概述:
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1. **文献名称**:*Escherichia coli DinB Homolog is a DNA Polymerase, pol IV, Regulated by the SOS Response*
**作者**:Kim, S.R., Matsui, K., Yamada, M., Gruz, P., & Nohmi, T.
**摘要**:该研究证实dinB编码的蛋白(Pol IV)是一种由SOS反应调控的DNA聚合酶,通过重组表达发现其具有跨损伤合成活性,尤其在苯并芘损伤DNA模板上表现出高错误倾向复制能力。
2. **文献名称**:*Crystal Structure of DinB Homolog, a DNA Polymerase Specialized in Lesion Bypass*
**作者**:Ling, H., Boudsocq, F., Woodgate, R., & Yang, W.
**摘要**:通过解析dinB重组蛋白的晶体结构,揭示了其活性位点适应大体积DNA损伤的分子机制,并阐明其作为Y家族聚合酶的独特催化特性。
3. **文献名称**:*Role of DinB in Error-Prone Replication and Adaptive Mutation in Escherichia coli*
**作者**:McKenzie, G.J., & Rosenberg, S.M.
**摘要**:研究利用重组DinB蛋白发现其在压力诱导的适应性突变中起关键作用,表明DinB的低保真度复制可能促进细菌基因组在应激条件下的快速进化。
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这些文献涵盖了dinB重组蛋白的功能、结构及生物学意义,适用于对DNA损伤修复或细菌应激反应机制的研究参考。
DinB, also known as DNA polymerase IV, is a Y-family DNA polymerase found in *Escherichia coli* and other bacteria. It plays a critical role in the bacterial SOS response, a stress-induced mechanism activated by DNA damage. Unlike high-fidelity replicative polymerases, DinB exhibits error-prone translesion synthesis (TLS) activity, enabling replication across damaged DNA templates that would otherwise stall replication forks. This process, while promoting survival under genotoxic stress, increases mutation rates—a trade-off termed the "SOS mutator effect."
Recombinant DinB protein is produced through genetic engineering, typically by cloning the *dinB* gene into expression vectors (e.g., plasmids) and expressing it in bacterial hosts like *E. coli*. Purification methods often involve affinity chromatography tags (e.g., His-tag) for efficient isolation. The recombinant protein retains its canonical TLS functions, including the ability to bypass bulky DNA lesions like N2-dG adducts, though it lacks proofreading exonuclease activity.
Studies on DinB have provided insights into TLS mechanisms, mutagenesis, and bacterial stress adaptation. Its error-prone nature makes it a model for understanding antibiotic-induced mutagenesis and bacterial evolution. In biotechnology, engineered variants of DinB are explored for synthetic biology applications, such as targeted mutagenesis or DNA synthesis under challenging conditions. Comparative analyses with eukaryotic Y-family polymerases (e.g., human Pol κ) also contribute to cancer research, particularly in contexts of chemotherapy resistance linked to TLS activity. Despite its prokaryotic origin, DinB remains a valuable tool for probing fundamental questions about DNA repair and genome instability.
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