BioVector? pRHA67 鼠李糖誘導(dǎo)型表達(dá)載體 / BioVector? pRHA67 Rhamnose-Inducible Expression Vector

通用定義 / General Definition:

BioVector? pRHA67 是一種利用 L-鼠李糖（L-rhamnose） 誘導(dǎo)系統(tǒng)的大腸桿菌表達(dá)載體。它采用了來源于大腸桿菌鼠李糖操縱子的 $rhaBAD$ 啟動(dòng)子。該系統(tǒng)的獨(dú)特優(yōu)勢(shì)在于其**“線性調(diào)控”**特性：蛋白表達(dá)水平與添加的鼠李糖濃度成正比，且在無誘導(dǎo)劑時(shí)具有極低的背景表達(dá)（漏表達(dá)）。相比于傳統(tǒng)的 IPTG 誘導(dǎo)系統(tǒng)，pRHA67 在表達(dá)毒性蛋白、優(yōu)化蛋白折疊以及實(shí)現(xiàn)多基因精準(zhǔn)共表達(dá)方面表現(xiàn)卓越。

BioVector? pRHA67 is an E. coli expression vector utilizing the L-rhamnose induction system. It features the $rhaBAD$ promoter derived from the E. coli rhamnose operon. The system's unique advantage lies in its "linear tunability": protein expression levels are proportional to the concentration of rhamnose added, with extremely low basal expression (leakiness) in the absence of the inducer. Compared to traditional IPTG-inducible systems, pRHA67 is superior for expressing toxic proteins, optimizing protein folding, and achieving precise co-expression of multiple genes.

BioVector? pRHA67 載體技術(shù)說明書 (Plasmid Datasheet)

中文版說明書

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? pRHA67 表達(dá)載體

• 載體類型： 大腸桿菌誘導(dǎo)型表達(dá)載體

• 啟動(dòng)子： BioVector? $P_{rhaBAD}$ (鼠李糖誘導(dǎo))

• 抗性基因： 氨芐青霉素 (Ampicillin)

• 復(fù)制子：$pBR322$ (中等拷貝)

• 調(diào)控基因：$rhaR$ 和 $rhaS$（內(nèi)置于載體，確保在不同菌株中的調(diào)控能力）

2. 培養(yǎng)與擴(kuò)增條件

• 宿主菌： BioVector? DH5$\alpha$（擴(kuò)增）；BL21 (DE3) 或其它常規(guī)表達(dá)菌株（表達(dá)）。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基（注意：避免使用含葡萄糖的培養(yǎng)基，除非為了進(jìn)一步抑制背景表達(dá)）。

• 抗生素： BioVector? 氨芐青霉素 (100 $\mu$g/mL)。

• 培養(yǎng)環(huán)境： 37°C 恒溫振蕩培養(yǎng)。

3. 載體應(yīng)用

• 毒性蛋白表達(dá)： 利用其嚴(yán)謹(jǐn)?shù)拈_關(guān)控制，防止蛋白在克隆過程中對(duì)宿主產(chǎn)生毒性。

• 蛋白濃度微調(diào)： 通過改變鼠李糖濃度（0.001% - 0.2%），精確尋找最佳的可溶性表達(dá)點(diǎn)。

• 多質(zhì)粒協(xié)同： 常與 IPTG 或阿拉伯糖誘導(dǎo)系統(tǒng)配合，實(shí)現(xiàn)不同組分的同時(shí)或先后誘導(dǎo)。

4. 注意事項(xiàng)

• 葡萄糖效應(yīng)： 葡萄糖通過分解代謝物阻遏效應(yīng)（Catabolite repression）抑制 $P_{rhaBAD}$。如需完全關(guān)閉表達(dá)，可在培養(yǎng)基中加入 0.2% 葡萄糖。

• 誘導(dǎo)劑用量： 常用誘導(dǎo)濃度為 0.1% - 0.2% $L$-鼠李糖。

English Datasheet

1. General Product Information

• Product Name: BioVector? pRHA67 Expression Vector

• Vector Type:E. coli Inducible Expression Vector

• Promoter: BioVector? $P_{rhaBAD}$ (Rhamnose-inducible)

• Antibiotic Resistance: Ampicillin

• Replicon:$pBR322$ (Medium copy)

• Regulatory Genes:$rhaR$ and $rhaS$ (Encoded on the vector for robust control across various strains)

2. Culture and Amplification Conditions

• Host Strains: BioVector? DH5$\alpha$ (for propagation); BL21 (DE3) or standard expression strains (for production).

• Culture Medium: BioVector? LB Medium (Note: Avoid glucose unless additional repression is required).

• Antibiotic Supplement: BioVector? Ampicillin (100 $\mu$g/mL).

• Incubation: 37°C with constant shaking.

3. Applications

• Toxic Protein Production: Utilizing tight transcriptional control to prevent host toxicity during growth phases.

• Tunable Expression: Adjusting rhamnose concentrations (0.001% to 0.2%) to find the optimal point for protein solubility and activity.

• Orthogonal Control: Compatible with IPTG or Arabinose systems for independent regulation of multiple pathways.

4. Key Usage Notes

• Glucose Inhibition: The $P_{rhaBAD}$ promoter is subject to catabolite repression by glucose. Adding 0.2% glucose to the medium can be used to ensure a completely "off" state.

• Inducer Concentration: Typical working concentrations range from 0.1% to 0.2% $L$-rhamnose.

Note: To maximize protein solubility, BioVector? recommends testing induction at lower temperatures (e.g., 20°C - 25°C) following the addition of rhamnose. For high-purity plasmid preparation, use the BioVector? Endotoxin-Free Plasmid Kit.

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