| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | polA |
| Uniprot No | P19821 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 293-832aa |
| 氨基酸序列 | ALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKALRDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEEAGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALSLEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAAVLEALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSSSDPNLQNIPVRTPLGQRIRRAFIAEEGWLLVALDYSQIELRVLAHLSGDENLIRVFQEGRDIHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERYFQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERKAFNMPVQGTAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYPLAVPLEVEVGIGEDWLSAKE |
| 预测分子量 | 61.0 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polA重组蛋白的3篇参考文献的简要概括：
1. **《Cloning and Expression of the polA Gene in Escherichia coli》**
- 作者：Kelley, W.S., et al.
- 摘要：该研究首次报道了大肠杆菌polA基因(编码DNA聚合酶I)的克隆与重组表达，验证了重组蛋白的切口平移活性，为后续酶学应用奠定基础。
2. **《Structural Analysis of DNA Polymerase I from E. coli》**
- 作者：Joyce, C.M., & Grindley, N.D.
- 摘要：通过X射线晶体学解析了重组polA蛋白的三维结构，阐明了其5'→3'聚合酶及5'→3'外切酶功能域的作用机制。
3. **《Optimization of Recombinant polA Expression for Molecular Biology Applications》**
- 作者：Smith, J., et al.
- 摘要：研究利用大肠杆菌表达系统优化重组polA蛋白的产量，通过诱导条件调整使酶活性提升3倍，并成功应用于高效DNA标记实验。
注：以上文献信息为示例性质，实际引用时需核实具体来源及细节。
**Background of polA Recombinant Protein**
The *polA* gene, encoding DNA polymerase I (Pol I), is a well-characterized enzyme originally identified in *Escherichia coli*. Discovered in the 1950s, Pol I plays a critical role in bacterial DNA replication and repair, functioning primarily in filling gaps during DNA synthesis, excising RNA primers, and participating in base excision repair. It possesses three distinct enzymatic activities: 5'→3' DNA polymerase activity, 3'→5' proofreading exonuclease activity, and 5'→3' exonuclease activity for nick translation.
Recombinant polA protein is produced through genetic engineering, where the *polA* gene is cloned into expression vectors and expressed in heterologous hosts like *E. coli*. This allows large-scale production of the enzyme with high purity and consistency. Historically, Pol I was pivotal in early molecular biology techniques, including cDNA synthesis and the first polymerase chain reaction (PCR) protocols, though it was later supplanted by thermostable polymerases like Taq.
Modern applications of recombinant Pol I leverage its nick-translation activity for DNA labeling and its role in *in vitro* DNA manipulation. A truncated version, the Klenow fragment (lacking 5'→3' exonuclease activity), is widely used in sequencing, mutagenesis, and blunt-end DNA preparation. Engineering polA variants with enhanced stability, altered processivity, or reduced exonuclease activity has expanded its utility in synthetic biology and diagnostics.
Research on polA recombinant protein continues to explore its structural and functional adaptability, providing insights into DNA metabolism and enabling tailored enzymatic tools for biotechnology. Its historical significance and versatility ensure its enduring relevance in both basic research and industrial applications.
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