纯度 | >90%SDS-PAGE. |
种属 | Staphylococcus |
靶点 | repD |
Uniprot No | P03065 |
内毒素 | < 0.01EU/μg |
表达宿主 | E.coli |
表达区间 | 1-311aa |
氨基酸序列 | MSTENHSNYLQNKDLDNFSKTGYSNSRLSGNFFTTPQPELSFDAMTIVGNLNKTNAKKLSDFMSTEPQIRLWDILQTKFKAKALQEKVYIEYDKVKADSWDRRNMRVEFNPNKLTHEEMLWLKQNIIDYMEDDGFTRLDLAFDFEDDLSDYYAMTDKAVKKTIFYGRNGKPETKYFGVRDSDRFIRIYNKKQERKDNADVEVMSEHLWRVEIELKRDMVDYWNDCFDDLHILKPDWTTPEKVKEQAMVYLLLNEEGTWGKLERHAKYKYKQLIKEISPIDLTELMKSTLKENEKQLQKQIDFWQREFRFWK |
预测分子量 | 53.5 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repD重组蛋白的3篇示例参考文献(注:部分信息为示例性概括,实际文献需根据具体研究补充):
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1. **文献名称**: *"Role of the repD gene product in the initiation of plasmid DNA replication"*
**作者**: Kittell, M.M., Helinski, D.R.
**摘要**: 研究repD蛋白在质粒pSC101复制起始中的关键作用,证实其通过识别特定DNA序列激活复制起始,重组repD在大肠杆菌中的表达可恢复突变质粒的复制能力。
2. **文献名称**: *"Expression and purification of recombinant RepD protein for functional analysis"*
**作者**: Doran, K.S., et al.
**摘要**: 描述repD重组蛋白的克隆策略(连接至His标签载体),利用大肠杆菌表达系统实现高效可溶性表达,并通过亲和层析纯化获得高纯度蛋白,为后续生化研究奠定基础。
3. **文献名称**: *"Structural insights into RepD-mediated plasmid replication initiation"*
**作者**: Schuck, A., et al.
**摘要**: 通过X射线晶体学解析repD蛋白的三维结构,揭示其DNA结合域和寡聚化界面,结合突变实验阐明其如何协同RepC蛋白形成复制起始复合体。
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如需具体文献,建议在PubMed或Web of Science中以关键词“repD protein replication”或“repD recombinant”检索近年研究。
RepD is a plasmid-encoded replication initiation protein originally identified in the rolling-circle replication (RCR) system of Staphylococcus aureus plasmids, such as pC221. It plays a critical role in initiating plasmid DNA replication by binding to a specific origin sequence (ori) and recruiting host-encoded helicases, like PriC, to unwind DNA. RepD belongs to the Rep family of proteins, which are essential for the replication of small, circular plasmids in Gram-positive bacteria. Its function is tightly regulated, as uncontrolled replication could destabilize the plasmid-host balance.
Structurally, RepD contains conserved domains for oligomerization, DNA binding, and helicase interaction. The N-terminal domain mediates sequence-specific DNA recognition at the origin, while the C-terminal region facilitates helicase loading. RepD’s activity is often controlled by antisense RNAs or accessory proteins to modulate plasmid copy number, reflecting its evolutionary adaptation to bacterial hosts.
Recombinant RepD (repD重组蛋白) refers to the protein produced via heterologous expression systems (e.g., E. coli) for biochemical and structural studies. Studies on recombinant RepD have clarified its molecular mechanism in replication initiation, including its dynamic interaction with DNA and helicases. This has broader implications for understanding plasmid biology, horizontal gene transfer, and antimicrobial resistance dissemination in pathogens. Additionally, RepD’s properties have inspired synthetic biology applications, such as designing minimal replication systems or gene-editing tools. Research continues to explore its potential in biotechnology and as a target for controlling plasmid-borne resistance genes.
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