| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | DHFR |
| Uniprot No | P00374 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 2-187aa |
| 氨基酸序列 | VGSLNCIVAVSQNMGIGKNGDLPWPPLRNEFRYFQRMTTTSSVEGKQNLVIMGKKTWFSIPEKNRPLKGRINLVLSRELKEPPQGAHFLSRSLDDALKLTEQPELANKVDMVWIVGGSSVYKEAMNHPGHLKLFVTRIMQDFESDTFFPEIDLEKYKLLPEYPGVLSDVQEEKGIKYKFEVYEKND |
| 预测分子量 | 37.3kDa |
| 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3篇关于DHFR(二氢叶酸还原酶)重组蛋白研究的代表性文献摘要信息，供参考：
1. **文献名称**：*High-yield expression and purification of recombinant human dihydrofolate reductase*
**作者**：Smith J, et al.
**摘要**：报道了利用大肠杆菌表达系统高效表达人源DHFR重组蛋白的优化方法，通过密码子优化和诱导条件调控，使蛋白表达量提高3倍，并采用亲和层析纯化获得高纯度酶，验证了其催化活性。
2. **文献名称**：*Crystal structure of a bifunctional dihydrofolate reductase-thymidylate synthase from Plasmodium falciparum*
**作者**：Brown NC, et al.
**摘要**：通过X射线晶体学解析了恶性疟原虫DHFR与胸苷酸合成酶的融合蛋白三维结构(分辨率2.8Å)，揭示了其双功能催化机制及抗疟药物结合位点，为抗寄生虫药物设计提供结构基础。
3. **文献名称**：*Engineering a thermostable dihydrofolate reductase for applications in nanotechnology*
**作者**：Garcia A, et al.
**摘要**：通过定向进化技术改造大肠杆菌DHFR，获得耐高温(75℃仍保持活性)突变体，并证明其可在纳米粒子组装中作为稳定连接元件，拓展了酶在生物材料领域的应用。
4. **文献名称**：*Comparative analysis of DHFR expression in eukaryotic vs prokaryotic systems*
**作者**：Johnson RL, et al.
**摘要**：系统比较了DHFR在酵母(毕赤酵母)和大肠杆菌中的表达差异，发现真核系统能实现正确折叠但产量较低，原核系统产量高但需重折叠，提出选择表达系统应根据下游应用需求。
注：以上为虚拟文献案例，实际研究需检索PubMed或Web of Science获取真实文献(如搜索关键词：recombinant DHFR protein expression)。经典文献可参考《Journal of Biological Chemistry》《Protein Expression and Purification》等期刊。
**Background of Recombinant Dihydrofolate Reductase (DHFR)**
Dihydrofolate reductase (DHFR) is a critical enzyme in cellular metabolism, catalyzing the reduction of dihydrofolate (DHF) to tetrahydrofolate (THF), a vital cofactor in nucleotide biosynthesis. This reaction is essential for DNA synthesis, amino acid metabolism, and cell proliferation. Due to its central role in cell growth, DHFR is a well-studied target for antibiotics, antifungals, and chemotherapeutic agents like methotrexate (MTX), which inhibit its activity to suppress rapidly dividing cells.
Recombinant DHFR refers to the protein produced through genetic engineering in heterologous expression systems, such as *E. coli*, yeast, or mammalian cells. The cloning and expression of the *dhfr* gene enable large-scale production of the enzyme for research and industrial applications. Recombinant DHFR retains the functional properties of the native enzyme, making it invaluable for biochemical studies, drug discovery, and structural biology. For example, X-ray crystallography studies using recombinant DHFR have elucidated its active site architecture and mechanism of action, guiding the design of targeted inhibitors.
In biotechnology, DHFR is also employed as a selection marker in mammalian cell culture systems. Cells lacking endogenous DHFR (e.g., CHO-DHFR⁻) require exogenous thymidine and glycine for survival. By introducing a recombinant *dhfr* gene alongside a gene of interest, researchers can select for high-producing cell lines using methotrexate, which forces gene amplification and enhances protein yield. This system is widely used in biopharmaceutical production, including monoclonal antibodies and therapeutic proteins.
Overall, recombinant DHFR bridges fundamental science and practical applications, serving as a model enzyme for understanding enzyme kinetics, a tool for bioproduction, and a cornerstone in anticancer and antimicrobial drug development. Its versatility continues to drive innovations across multiple disciplines.
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