BioVector® Mito-R-GECO1 线粒体红色荧光钙离子指示质粒 / Mito-R-GECO1 Mitochondrial Red Calcium Indicator Plasmid

通用定义 / General Definition:

BioVector® Mito-R-GECO1 是一种专用于监测线粒体内钙离子 ($Ca^{2+}$) 动态的基因编码钙指示剂 (GECI)。该质粒表达一种红色荧光蛋白（基于 R-GECO1），并通过在 $N$ 端融合线粒体靶向序列（通常来源于 $Cytochrome$$c$$oxidase$$subunit$$VIII$, $COX8$），将指示剂精确锁定在线粒体基质中。当线粒体内 $Ca^{2+}$ 浓度升高时，其红色荧光强度会随之增加，是研究线粒体代谢、凋亡及细胞信号转导的强有力工具。

BioVector® Mito-R-GECO1 is a genetically encoded calcium indicator (GECI) specifically designed for monitoring mitochondrial calcium ($Ca^{2+}$) dynamics. The plasmid expresses a red fluorescent protein (based on R-GECO1) fused to a mitochondrial targeting sequence (typically from $Cytochrome$$c$$oxidase$$subunit$$VIII$, $COX8$). This ensures the indicator is precisely localized within the mitochondrial matrix. Upon an increase in mitochondrial $Ca^{2+}$ concentration, the red fluorescence intensity increases, making it a powerful tool for studying mitochondrial metabolism, apoptosis, and cellular signaling.

BioVector® Mito-R-GECO1 技术说明书 (Plasmid Datasheet)

中文版说明书

1. 产品基本信息

• 产品名称： BioVector® Mito-R-GECO1 钙指示质粒

• 指示剂类型： 强度型红色荧光钙指示剂 (R-GECO1)

• 靶向定位： 线粒体基质 (Mitochondrial Matrix)

• 启动子： BioVector® CMV 强启动子

• 抗性基因： * 大肠杆菌： kanamycin (卡那霉素) 或 Ampicillin (氨苄青霉素)

  • 真核筛选： Neomycin (G418) 或 无（取决于骨架）

• 激发/发射波长：$Ex/Em = 561 / 589$ nm

2. 培养与扩增条件

• 宿主菌： BioVector® DH5$\alpha$ 或 TOP10。

• 培养基： BioVector® LB 培养基。

• 抗生素： 根据载体要求添加 (如 50 $\mu$g/mL 卡那霉素)。

• 培养环境： 37°C 恒温振荡培养。

3. 载体应用

• 线粒体功能分析： 实时监测刺激条件下线粒体对 $Ca^{2+}$ 的吸收与释放。

• 多色成像： 与绿色荧光指示剂（如 GCaMP）配合，同时观察细胞质和线粒体 $Ca^{2+}$。

• 药物筛选： 评估药物对线粒体钙稳态的影响。

4. 注意事项

• 定位验证： 转染后建议使用 BioVector® MitoTracker 对细胞进行共染，验证 R-GECO1 的线粒体定位准确性。

• 光漂白： 红色荧光蛋白虽较稳定，但长时间高强度激发仍可能发生光漂白，建议优化成像参数。

English Datasheet

1. General Product Information

• Product Name: BioVector® Mito-R-GECO1 Calcium Indicator Plasmid

• Indicator Type: Intensity-based Red Fluorescent Calcium Indicator (R-GECO1)

• Localization: Mitochondrial Matrix

• Promoter: BioVector® CMV constitutive promoter

• Selection Markers: * E. coli: Kanamycin or Ampicillin

  • Eukaryotic: Neomycin (G418) or None (depending on backbone)

• Excitation/Emission:$Ex/Em = 561 / 589$ nm

2. Culture and Amplification Conditions

• Host Strains: BioVector® DH5$\alpha$ or TOP10.

• Culture Medium: BioVector® LB Medium.

• Antibiotic Supplement: As per vector requirement (e.g., 50 $\mu$g/mL Kanamycin).

• Incubation: 37°C with constant shaking.

3. Applications

• Mitochondrial Function Assays: Real-time monitoring of mitochondrial $Ca^{2+}$ uptake and release under various stimuli.

• Multi-color Imaging: Co-imaging with green indicators (e.g., GCaMP) to observe cytosolic and mitochondrial $Ca^{2+}$ simultaneously.

• Drug Discovery: Evaluating the effects of compounds on mitochondrial calcium homeostasis.

4. Key Usage Notes

• Localization Verification: Post-transfection, it is recommended to co-stain with BioVector® MitoTracker to confirm the accurate mitochondrial localization of the R-GECO1 signal.

• Photobleaching: While R-GECO1 is relatively stable, prolonged exposure to high-intensity excitation light can cause photobleaching; optimization of imaging frequency and intensity is advised.

Note: For highest imaging quality, use BioVector® phenol red-free medium during live-cell imaging to reduce background fluorescence. Always ensure plasmids are extracted using BioVector® endotoxin-free kits for sensitive primary cell transfections.

Supplier生产厂家: BioVector NTCC典型培养物保藏中心

E-mail: BioVector@163.com

http://www.biovector.net