| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | rnhB |
| Uniprot No | O29634 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-205aa |
| 氨基酸序列 | MKAGIDEAGKGCVIGPLVVAGVACSDEDRLRKLGVKDSKKLSQGRREELAEEIRKICRTEVLKVSPENLDERMAAKTINEILKECYAEIILRLKPEIAYVDSPDVIPERLSRELEEITGLRVVAEHKADEKYPLVAAASIIAKVEREREIERLKEKFGDFGSGYASDPRTREVLKEWIASGRIPSCVRMRWKTVSNLRQKTLDDF |
| 预测分子量 | 23,1 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rnhB重组蛋白的3篇代表性文献示例(注：部分文献为模拟概括，实际引用需核实)：
1. **"Characterization of Recombinant RNase HII from Escherichia coli: Role in DNA Repair"**
*作者：Yokoyama et al. (2005)*
摘要：研究通过大肠杆菌表达系统纯化rnhB编码的RNase HII重组蛋白，分析其酶学特性及在DNA错配修复中的功能，证明其可特异性降解RNA-DNA杂交链中的RNA部分。
2. **"Thermostable Recombinant RNase HII from Thermococcus kodakarensis: Expression and Biotechnological Applications"**
*作者：Chen et al. (2012)*
摘要：报道了嗜热古菌Thermococcus kodakarensis来源的rnhB基因在大肠杆菌中的重组表达，纯化获得高热稳定性的RNase HII，并验证其在PCR技术中减少非特异性扩增的应用潜力。
3. **"Structural Insights into the Catalytic Mechanism of Recombinant RNase HII from Bacillus subtilis"**
*作者：Zhang et al. (2018)*
摘要：通过X射线晶体学解析枯草芽孢杆菌RNase HII重组蛋白的三维结构，揭示其活性中心关键氨基酸残基的作用，为设计新型RNA-DNA杂交链靶向工具提供依据。
4. **"CRISPR-Cas9 Genome Editing Enhancement by Recombinant RNase HII in vitro"**
*作者：Smith et al. (2021)*
摘要：研究利用重组表达的RNase HII与CRISPR系统联用，证明其可有效降解向导RNA与DNA的非特异性结合，显著降低基因编辑脱靶效应。
(注：以上文献标题及内容为示例性模拟，具体研究请以实际数据库检索结果为准。)
**Background of RnhB Recombinant Protein**
RNase HII (encoded by the *rnhB* gene) is a member of the ribonuclease H (RNase H) family, which specifically cleaves RNA strands in RNA-DNA hybrid substrates. This enzymatic activity is critical for maintaining genome stability by resolving R-loops—three-stranded nucleic acid structures formed during transcription or replication—and repairing misincorporated ribonucleotides in DNA. Unlike RNase HI (encoded by *rnhA*), which is found predominantly in bacteria and eukaryotes, RNase HII is evolutionarily conserved across all domains of life, including archaea, bacteria, and eukaryotes. Its substrate specificity and dependence on divalent metal ions (e.g., Mg²⁺ or Mn²⁺) for catalytic activity distinguish it from other RNase H subtypes.
The recombinant RnhB protein is produced through heterologous expression, typically in *Escherichia coli*, by cloning the *rnhB* gene into an expression vector under an inducible promoter. This approach enables large-scale production of the enzyme for biochemical and structural studies. Recombinant RnhB is purified using affinity chromatography, often leveraging tags such as polyhistidine (His-tag) for efficient isolation. Its stability, solubility, and enzymatic activity are optimized through buffer conditioning and removal of endotoxins or host contaminants.
Research on RnhB has revealed its role in DNA repair pathways, particularly in conjunction with polymerase-proofreading systems. It also serves as a model enzyme for studying RNase H mechanisms, including substrate recognition and metal ion coordination. In biotechnology, recombinant RnhB is utilized as a tool enzyme for manipulating RNA-DNA hybrids in molecular cloning and sequencing applications. Additionally, its conserved structure and function make it a potential target for antimicrobial drug development, especially in pathogens reliant on RNase HII activity for survival.
Overall, the study of recombinant RnhB provides insights into nucleic acid metabolism and supports applications ranging from basic enzymology to therapeutic innovation.
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