BioVector? pSAT1-cCFP-C (pE3242) 植物雙分子熒光互補(bǔ)載體 / pSAT1-cCFP-C (pE3242) Plant BiFC Vector

通用定義 / General Definition:BioVector? pSAT1-cCFP-C (pE3242) 是一種專門用于植物雙分子熒光互補(bǔ) (BiFC) 實(shí)驗(yàn)的衛(wèi)星載體。該載體包含青色熒光蛋白 (CFP) 的 C 端片段 (cCFP)。在實(shí)驗(yàn)中，將一個目的蛋白與 cCFP 融合，另一個候選蛋白與 nCFP（N 端片段）融合。如果這兩個蛋白在植物細(xì)胞內(nèi)發(fā)生相互作用，cCFP 和 nCFP 片段將在空間上靠近并重新折疊成完整的、具有功能的熒光蛋白，從而發(fā)出青色熒光。作為 pSAT 系列成員，它具有高度模塊化特征，便于通過罕見酶切位點(diǎn)轉(zhuǎn)移至二元表達(dá)系統(tǒng)。

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) 技術(shù)說明書 (Technical Datasheet)

中文版說明書

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? pSAT1-cCFP-C 表達(dá)載體

• 載體編號： pE3242

• 啟動子： 2x35S 啟動子（增強(qiáng)型花椰菜花葉病毒 35S 啟動子）

• 熒光片段： cCFP（CFP 的 C 端片段，C 端融合）

• 抗性基因： 氨芐青霉素 (Ampicillin)

• 終止子： TL 終止子

2. 培養(yǎng)與擴(kuò)增條件

• 宿主菌： BioVector? DH5-alpha 或 TOP10。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基。

• 抗生素： 100 ug/mL 氨芐青霉素。

• 環(huán)境： 37 攝氏度，恒溫振蕩。

3. 載體應(yīng)用

• 蛋白質(zhì)相互作用 (PPI)： 在擬南芥原生質(zhì)體、煙草葉片或穩(wěn)定轉(zhuǎn)基因植物中驗(yàn)證蛋白結(jié)合。

• 復(fù)合物定位： 確定蛋白質(zhì)復(fù)合體在細(xì)胞內(nèi)的具體分布位置。

• 信號通路研究： 監(jiān)測在特定刺激下蛋白質(zhì)相互作用的動態(tài)變化。

4. 注意事項(xiàng)

• 讀碼框一致性： 插入的目的基因必須移除終止密碼子，并確保與下文的 cCFP 片段保持正確的讀碼框?qū)R。

• 對照設(shè)置： 實(shí)驗(yàn)中必須包含空載體對照以及已知的陽性/陰性相互作用對照，以排除非特異性組裝。

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生產(chǎn)廠家: BioVector NTCC典型培養(yǎng)物保藏中心

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