BioVector? 35S-SPYCE(MR) 植物雙分子熒光互補(bǔ)載體 / 35S-SPYCE(MR) Plant BiFC Vector

通用定義 / General Definition:BioVector? 35S-SPYCE(MR) 是一種用于植物雙分子熒光互補(bǔ) (BiFC) 實(shí)驗(yàn)的表達(dá)載體。其名稱中的 SPYCE 代表 Split Potential Yellow Close Enhanced（即 YFP 的分割片段），而 (MR) 表示該載體攜帶 Modified Red (mRFP) 或類似的紅色熒光標(biāo)記作為內(nèi)參，或者在某些版本中代表特定的克隆位點(diǎn)排列。該載體含有增強(qiáng)型黃色熒光蛋白 (YFP) 的 C 端片段 (cYFP)。當(dāng)與含有 nYFP（N 端片段）的載體共轉(zhuǎn)染時(shí)，如果融合的兩個(gè)目的蛋白發(fā)生相互作用，則會(huì)重新組裝出完整的 YFP 熒光。35S 啟動(dòng)子確保了該系統(tǒng)在大多數(shù)植物組織中的強(qiáng)力表達(dá)。

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) 技術(shù)說(shuō)明書(shū) (Technical Datasheet)

中文版說(shuō)明書(shū)

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? 35S-SPYCE(MR) 表達(dá)載體

• 啟動(dòng)子： CaMV 35S 啟動(dòng)子

• 熒光片段： cYFP (YFP 的 C 端片段)

• 抗性基因： * 大腸桿菌： 卡那霉素 (Kanamycin) 或 氨芐青霉素 (取決于骨架版本)

  • 植物篩選： 通常含有潮霉素 (Hygromycin) 或 除草劑抗性 (Basta)

• 復(fù)制子： pUC 來(lái)源 (高拷貝)

2. 培養(yǎng)與擴(kuò)增條件

• 宿主菌： BioVector? DH5-alpha。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基。

• 環(huán)境： 37 攝氏度，恒溫振蕩。

3. 載體應(yīng)用

• 蛋白質(zhì)相互作用驗(yàn)證： 在煙草葉片瞬時(shí)表達(dá)或擬南芥原生質(zhì)體中檢測(cè)蛋白結(jié)合。

• 亞細(xì)胞定位： 觀察蛋白質(zhì)復(fù)合物在細(xì)胞核、細(xì)胞質(zhì)或細(xì)胞膜上的具體定位。

• 大規(guī)模篩選： 配合文庫(kù)載體進(jìn)行蛋白互作組學(xué)研究。

4. 注意事項(xiàng)

• 融合方向： SPYCE 載體通常用于目的蛋白的 C 端融合，請(qǐng)檢查克隆位點(diǎn)以確保讀碼框正確。

• 背景熒光： YFP 片段在沒(méi)有互作的情況下也可能發(fā)生微弱的非特異性自組裝，實(shí)驗(yàn)中必須設(shè)置空載體及陰性對(duì)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.

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