BioVector® 35S-SPYCE(MR) 植物双分子荧光互补载体 / 35S-SPYCE(MR) Plant BiFC Vector

通用定义 / General Definition:BioVector® 35S-SPYCE(MR) 是一种用于植物双分子荧光互补 (BiFC) 实验的表达载体。其名称中的 SPYCE 代表 Split Potential Yellow Close Enhanced（即 YFP 的分割片段），而 (MR) 表示该载体携带 Modified Red (mRFP) 或类似的红色荧光标记作为内参，或者在某些版本中代表特定的克隆位点排列。该载体含有增强型黄色荧光蛋白 (YFP) 的 C 端片段 (cYFP)。当与含有 nYFP（N 端片段）的载体共转染时，如果融合的两个目的蛋白发生相互作用，则会重新组装出完整的 YFP 荧光。35S 启动子确保了该系统在大多数植物组织中的强力表达。

BioVector® 35S-SPYCE(MR) is an expression vector designed for Bimolecular Fluorescence Complementation (BiFC) assays in plant systems. SPYCE refers to the split fragments of an enhanced yellow fluorescent protein (YFP). This specific vector contains the C-terminal fragment of YFP (cYFP). When co-transfected with a complementary nYFP vector, the interaction between two fused target proteins brings the YFP fragments together to reconstitute a functional fluorescent protein. The 35S promoter drives strong, constitutive expression across most plant tissues, making it a robust tool for protein-protein interaction (PPI) studies.

BioVector® 35S-SPYCE(MR) 技术说明书 (Technical Datasheet)

中文版说明书

1. 产品基本信息

• 产品名称： BioVector® 35S-SPYCE(MR) 表达载体

• 启动子： CaMV 35S 启动子

• 荧光片段： cYFP (YFP 的 C 端片段)

• 抗性基因： * 大肠杆菌： 卡那霉素 (Kanamycin) 或 氨苄青霉素 (取决于骨架版本)

  • 植物筛选： 通常含有潮霉素 (Hygromycin) 或 除草剂抗性 (Basta)

• 复制子： pUC 来源 (高拷贝)

2. 培养与扩增条件

• 宿主菌： BioVector® DH5-alpha。

• 培养基： BioVector® LB 培养基。

• 环境： 37 摄氏度，恒温振荡。

3. 载体应用

• 蛋白质相互作用验证： 在烟草叶片瞬时表达或拟南芥原生质体中检测蛋白结合。

• 亚细胞定位： 观察蛋白质复合物在细胞核、细胞质或细胞膜上的具体定位。

• 大规模筛选： 配合文库载体进行蛋白互作组学研究。

4. 注意事项

• 融合方向： SPYCE 载体通常用于目的蛋白的 C 端融合，请检查克隆位点以确保读码框正确。

• 背景荧光： YFP 片段在没有互作的情况下也可能发生微弱的非特异性自组装，实验中必须设置空载体及阴性对照。

English Datasheet

1. General Product Information

• Product Name: BioVector® 35S-SPYCE(MR) Expression Vector

• Promoter: CaMV 35S Promoter

• Fluorescence Fragment: cYFP (C-terminal fragment of YFP)

• Selection Markers: * E. coli: Kanamycin or Ampicillin (variant dependent)

  • Plant: Usually Hygromycin or Basta resistance

• Replicon: pUC origin (High-copy)

2. Culture and Amplification Conditions

• Host Strains: BioVector® DH5-alpha.

• Culture Medium: BioVector® LB Medium.

• Incubation: 37 degrees Celsius with constant shaking.

3. Applications

• Protein-Protein Interaction (PPI) Validation: Detecting protein binding in tobacco leaf transient assays or Arabidopsis protoplasts.

• Subcellular Localization: Identifying the specific site (nucleus, cytoplasm, or plasma membrane) where protein complexes reside.

• High-Throughput Screening: Used in conjunction with library vectors for interactome studies.

4. Key Usage Notes

• Fusion Orientation: SPYCE vectors are typically designed for C-terminal fusion of the target protein. Always verify the junction sequence to ensure the open reading frame (ORF) is maintained.

• Non-specific Complementation: Since split fluorescent fragments have a natural affinity for each other, it is essential to include negative controls (e.g., non-interacting proteins) to distinguish true biological interactions from background noise.

Note: For multi-vector transformations, BioVector® recommends verifying the antibiotic resistance compatibility of all components. To ensure high expression levels in plant tissues, use BioVector® Endotoxin-Free Plasmid Extraction Kits for all transient expression protocols.

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