BioVector® pRHA67 鼠李糖诱导型表达载体 / BioVector® pRHA67 Rhamnose-Inducible Expression Vector

通用定义 / General Definition:

BioVector® pRHA67 是一种利用 L-鼠李糖（L-rhamnose） 诱导系统的大肠杆菌表达载体。它采用了来源于大肠杆菌鼠李糖操纵子的 $rhaBAD$ 启动子。该系统的独特优势在于其**“线性调控”**特性：蛋白表达水平与添加的鼠李糖浓度成正比，且在无诱导剂时具有极低的背景表达（漏表达）。相比于传统的 IPTG 诱导系统，pRHA67 在表达毒性蛋白、优化蛋白折叠以及实现多基因精准共表达方面表现卓越。

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 载体技术说明书 (Plasmid Datasheet)

中文版说明书

1. 产品基本信息

• 产品名称： BioVector® pRHA67 表达载体

• 载体类型： 大肠杆菌诱导型表达载体

• 启动子： BioVector® $P_{rhaBAD}$ (鼠李糖诱导)

• 抗性基因： 氨苄青霉素 (Ampicillin)

• 复制子：$pBR322$ (中等拷贝)

• 调控基因：$rhaR$ 和 $rhaS$（内置于载体，确保在不同菌株中的调控能力）

2. 培养与扩增条件

• 宿主菌： BioVector® DH5$\alpha$（扩增）；BL21 (DE3) 或其它常规表达菌株（表达）。

• 培养基： BioVector® LB 培养基（注意：避免使用含葡萄糖的培养基，除非为了进一步抑制背景表达）。

• 抗生素： BioVector® 氨苄青霉素 (100 $\mu$g/mL)。

• 培养环境： 37°C 恒温振荡培养。

3. 载体应用

• 毒性蛋白表达： 利用其严谨的开关控制，防止蛋白在克隆过程中对宿主产生毒性。

• 蛋白浓度微调： 通过改变鼠李糖浓度（0.001% - 0.2%），精确寻找最佳的可溶性表达点。

• 多质粒协同： 常与 IPTG 或阿拉伯糖诱导系统配合，实现不同组分的同时或先后诱导。

4. 注意事项

• 葡萄糖效应： 葡萄糖通过分解代谢物阻遏效应（Catabolite repression）抑制 $P_{rhaBAD}$。如需完全关闭表达，可在培养基中加入 0.2% 葡萄糖。

• 诱导剂用量： 常用诱导浓度为 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.

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