BioVector® pSAT1-cCFP-C (pE3242) 植物双分子荧光互补载体 / pSAT1-cCFP-C (pE3242) Plant BiFC Vector

通用定义 / General Definition:BioVector® pSAT1-cCFP-C (pE3242) 是一种专门用于植物双分子荧光互补 (BiFC) 实验的卫星载体。该载体包含青色荧光蛋白 (CFP) 的 C 端片段 (cCFP)。在实验中，将一个目的蛋白与 cCFP 融合，另一个候选蛋白与 nCFP（N 端片段）融合。如果这两个蛋白在植物细胞内发生相互作用，cCFP 和 nCFP 片段将在空间上靠近并重新折叠成完整的、具有功能的荧光蛋白，从而发出青色荧光。作为 pSAT 系列成员，它具有高度模块化特征，便于通过罕见酶切位点转移至二元表达系统。

BioVector® pSAT1-cCFP-C (pE3242) is a satellite vector specifically designed for Bimolecular Fluorescence Complementation (BiFC) assays in plant systems. This vector contains the C-terminal fragment of Cyan Fluorescent Protein (cCFP). In a typical experiment, one target protein is fused to cCFP, while another candidate protein is fused to nCFP (the N-terminal fragment). If these two proteins interact within the plant cell, the cCFP and nCFP fragments are brought into close proximity and reconstitute into a functional fluorescent protein, emitting cyan light. As a member of the pSAT series, it is highly modular, allowing for easy transfer to binary expression systems via rare-cutting restriction sites.

BioVector® pSAT1-cCFP-C (pE3242) 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® pSAT1-cCFP-C 表达载体

• 载体编号： pE3242

• 启动子： 2x35S 启动子（增强型花椰菜花叶病毒 35S 启动子）

• 荧光片段： cCFP（CFP 的 C 端片段，C 端融合）

• 抗性基因： 氨苄青霉素 (Ampicillin)

• 终止子： TL 终止子

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： 100 ug/mL 氨苄青霉素。

• 环境： 37 摄氏度，恒温振荡。

3. 载体应用

• 蛋白质相互作用 (PPI)： 在拟南芥原生质体、烟草叶片或稳定转基因植物中验证蛋白结合。

• 复合物定位： 确定蛋白质复合体在细胞内的具体分布位置。

• 信号通路研究： 监测在特定刺激下蛋白质相互作用的动态变化。

4. 注意事项

• 读码框一致性： 插入的目的基因必须移除终止密码子，并确保与下文的 cCFP 片段保持正确的读码框对齐。

• 对照设置： 实验中必须包含空载体对照以及已知的阳性/阴性相互作用对照，以排除非特异性组装。

English Datasheet

1. General Product Information

• Product Name: BioVector® pSAT1-cCFP-C Expression Vector

• Vector ID: pE3242

• Promoter: Double 35S Promoter (2x35S CaMV)

• Fluorescence Fragment: cCFP (C-terminal fragment of CFP, C-terminal fusion)

• Antibiotic Resistance: Ampicillin (100 ug/mL)

• Terminator: TL Terminator

2. Culture and Amplification Conditions

• Host Strains: BioVector® DH5-alpha or TOP10.

• Culture Medium: BioVector® LB Medium.

• Incubation: 37 degrees Celsius with constant shaking.

3. Applications

• Protein-Protein Interactions (PPI): Validating protein binding in Arabidopsis protoplasts, tobacco leaves, or stable transgenic plants.

• Complex Localization: Identifying the specific subcellular site where a protein complex assembles.

• Signaling Pathways: Monitoring dynamic changes in protein associations under specific physiological stimuli.

4. Key Usage Notes

• Reading Frame: Ensure the Gene of Interest (GOI) lacks a stop codon and is cloned in-frame with the downstream cCFP fragment.

• Experimental Controls: It is essential to include empty vector controls and known positive/negative interaction pairs to account for potential non-specific protein self-assembly.

Note: For multi-protein studies, pSAT1-cCFP-C is often paired with nCFP-tagged vectors. BioVector® recommends using the pPZP-RCS modular binary vector system for Agrobacterium-mediated co-transformation to ensure stable and high-level expression of both BiFC components.

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