纯度 | >90%SDS-PAGE. |
种属 | Human |
靶点 | MGMT |
Uniprot No | P16455 |
内毒素 | < 0.01EU/μg |
表达宿主 | E.coli |
表达区间 | 1-207aa |
氨基酸序列 | MDKDCEMKRT TLDSPLGKLE LSGCEQGLHE IKLLGKGTSA ADAVEVPAPA AVLGGPEPLM QCTAWLNAYF HQPEAIEEFP VPALHHPVFQ QESFTRQVLW KLLKVVKFGE VISYQQLAAL AGNPKAARAV GGAMRGNPVP ILIPCHRVVC SSGAVGNYSG GLAVKEWLLA HEGHRLGKPG LGGSSGLAGA WLKGAGATSG SPPAGRN |
预测分子量 | 25 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3篇关于MGMT重组蛋白的示例文献(注:内容为示例性概括,非真实文献):
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1. **标题**: "Expression and purification of recombinant human O6-methylguanine-DNA methyltransferase (MGMT)"
**作者**: Smith A, et al.
**摘要**: 研究报道了在大肠杆菌中高效表达人源MGMT重组蛋白的优化方法,通过His标签亲和层析纯化,验证了其修复DNA烷基化损伤的酶活性,为体外药物筛选提供基础。
2. **标题**: "Structural insights into MGMT-mediated DNA repair by crystal analysis"
**作者**: Jones B, et al.
**摘要**: 通过X射线晶体学解析重组MGMT蛋白的三维结构,揭示了其活性位点与O6-甲基鸟嘌呤的特异性结合机制,为设计MGMT抑制剂增强化疗效果提供理论依据。
3. **标题**: "MGMT overexpression confers temozolomide resistance in glioblastoma models"
**作者**: Lee C, et al.
**摘要**: 利用重组MGMT蛋白转导胶质瘤细胞,证明其过表达显著降低替莫唑胺(TMZ)的细胞毒性,提示MGMT活性是预测化疗耐药的关键生物标志物。
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(如需真实文献,建议通过PubMed或Web of Science以关键词"recombinant MGMT protein"检索。)
**Background of MGMT Recombinant Protein**
MGMT (O⁶-methylguanine-DNA methyltransferase) is a DNA repair enzyme critical for maintaining genomic stability by reversing alkylation damage at the O⁶ position of guanine. This specific lesion, often induced by alkylating agents (e.g., temozolomide or nitrosoureas), disrupts DNA replication and triggers cytotoxic or mutagenic effects. MGMT functions as a "suicide enzyme," transferring the alkyl group from damaged DNA to its active-site cysteine residue, thereby irreversibly inactivating itself. This repair mechanism is pivotal in protecting cells from alkylation-induced toxicity but also contributes to resistance in cancer chemotherapy, as elevated MGMT levels in tumors diminish the efficacy of alkylating drugs.
In cancer biology, MGMT expression is epigenetically regulated, with promoter methylation silencing the gene in certain tumors. This methylation status is a predictive biomarker; for example, glioblastoma patients with MGMT-silenced tumors show better responses to temozolomide. Recombinant MGMT protein, produced via bacterial (e.g., *E. coli*) or eukaryotic expression systems, is widely used in research to elucidate DNA repair mechanisms, screen for MGMT inhibitors, or model resistance pathways. Its applications extend to gene therapy, where engineered MGMT variants (e.g., P140K mutant) are employed to protect hematopoietic stem cells during dose-intensified chemotherapy.
The development of MGMT recombinant tools has advanced personalized oncology and genome-editing studies, offering insights into tailoring chemotherapeutic regimens and improving therapeutic outcomes. However, challenges remain in balancing DNA repair modulation to avoid unintended mutagenesis or therapeutic resistance. Ongoing research focuses on optimizing MGMT-based strategies to enhance precision in cancer treatment and reduce off-target effects.
在生物科技领域,蛋白研发与生产是前沿探索的关键支撑。艾普蒂作为行业内的创新者,凭借自身卓越的研发实力,每年能成功研发 1000 多种全新蛋白,在重组蛋白领域不断突破。 在重组蛋白生产过程中,艾普蒂积累了丰富且成熟的经验。从结构复杂的跨膜蛋白,到具有特定催化功能的酶、参与信号传导的激酶,再到用于免疫研究的病毒抗原,艾普蒂都能实现高效且稳定的生产。 这一成就离不开艾普蒂强大的技术平台。我们构建了多元化的重组蛋白表达系统,昆虫细胞、哺乳动物细胞以及原核蛋白表达系统协同运作。不同的表达系统各有优势,能够满足不同客户对重组蛋白的活性、产量、成本等多样化的需求,从而提供高品质、低成本的活性重组蛋白。 艾普蒂提供的不只是产品,更是从源头到终端的一站式解决方案。从最初的基因合成,精准地构建出符合要求的基因序列,到载体构建,为蛋白表达创造适宜的环境,再到蛋白质表达和纯化,每一个环节都严格把控。我们充分尊重客户的个性化需求,在表达 / 纯化标签的选择、表达宿主的确定等方面,为客户量身定制专属方案。 同时,艾普蒂还配备了多种纯化体系,能够应对不同特性蛋白的纯化需求。这种灵活性和专业性,极大地提高了蛋白表达和纯化的成功率,让客户的研究项目得以顺利推进,在生物科技的探索道路上助力每一位科研工作者迈向成功。
艾普蒂生物自主研发并建立综合性重组蛋白生产和抗体开发技术平台,包括: 哺乳动物细胞表达平台:利用哺乳动物细胞精准修饰蛋白,产出与天然蛋白相似的重组蛋白,用于药物研发、细胞治疗等。 杂交瘤开发平台:通过细胞融合筛选出稳定分泌单克隆抗体的杂交瘤细胞株,优化后的技术让抗体亲和力与特异性更高,应用于疾病诊断、免疫治疗等领域。 单 B 细胞筛选平台:FACS 用荧光标记和流式细胞仪快速分选特定 B 细胞;Beacon® 基于微流控技术,单细胞水平捕获、分析 B 细胞,挖掘抗体多样性,缩短开发周期。 凭借这些平台,艾普蒂生物为客户提供优质试剂和专业 CRO 技术服务,推动生物科技发展。
艾普蒂生物在重组蛋白和天然蛋白开发领域经验十分丰富,拥有超过 2 万种重组蛋白的开发案例。在四大重组蛋白表达平台的运用上,艾普蒂生物不仅经验老到,还积累了详实的成功案例。针对客户的工业化生产需求,我们能够定制并优化实验方案。通过小试探索、工艺放大以及条件优化等环节,对重组蛋白基因序列进行优化,全面探索多种条件,精准找出最契合客户需求的生产方法。 此外,公司还配备了自有下游验证平台,可对重组蛋白展开系统的质量检测与性能测试,涵盖蛋白互作检测、活性验证、内毒素验证等,全方位保障产品质量。 卡梅德生物同样重视蛋白工艺开发,确保生产出的蛋白质具备所需的纯度、稳定性与生物活性,这对于保障药物的安全性和有效性起着关键作用 ,与艾普蒂生物共同推动着行业的发展。
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