BioVector® pSAM-Bt 苏云金芽孢杆菌穿梭表达质粒载体 / BioVector® pSAM-Bt Plasmid Vector

通用定义 / General Definition:BioVector® pSAM-Bt 是一种专门用于苏云金芽孢杆菌 (Bacillus thuringiensis, Bt) 的穿梭质粒或表达载体。该载体通常包含在大肠杆菌 (E. coli) 和芽孢杆菌中均能发挥作用的复制起始位点，使其能够在这两种宿主之间进行基因穿梭。它被广泛应用于 Bt 杀虫晶体蛋白（Cry 蛋白）的异源表达、启动子功能表征以及针对农业害虫的基因工程菌株改良。

BioVector® pSAM-Bt is a specialized shuttle or expression vector designed for use in Bacillus thuringiensis (Bt). The vector typically contains origins of replication functional in both E. coli and Bacillus species, facilitating gene shuttling between these two hosts. It is widely used for the heterologous expression of Bt insecticidal crystal proteins (Cry proteins), promoter characterization, and the genetic engineering of Bt strains for agricultural pest control.

BioVector® pSAM-Bt 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® pSAM-Bt 表达载体

• 载体类型： 芽孢杆菌穿梭质粒

• 常用宿主： 大肠杆菌 (E. coli)、苏云金芽孢杆菌 (B. thuringiensis)

• 抗性基因： * 大肠杆菌： 氨苄青霉素 (Ampicillin) 或 红霉素 (Erythromycin)

  • 芽孢杆菌： 红霉素 (Erythromycin) 或 氯霉素 (Chloramphenicol)（视具体版本而定）

• 复制子： pMB1 (E. coli); pBT1 或类似 (Bt)

2. 培养与扩增条件

• 大肠杆菌扩增： 使用 BioVector® LB 培养基，添加相应抗生素，37°C 培养。

• Bt 转化与筛选： 使用 BioVector® GYS 或 LB 培养基，添加 BioVector® 红霉素，通常在 28°C - 30°C 下培养以保持质粒稳定性。

• 诱导条件： 若带有特定启动子，需根据其特性（如胞外蛋白分泌或芽孢形成期表达）进行诱导。

3. 载体应用

• Bt 基因工程： 用于表达 Cry、Vip 等杀虫蛋白，增强菌株的杀虫谱。

• 启动子活性分析： 研究 Bt 芽孢形成阶段特异性启动子的调控机制。

• 蛋白质组学研究： 在 Bt 内源环境下生产具有天然活性的蛋白质。

4. 注意事项

• 质粒稳定性： 在 Bt 中培养时，由于质粒拷贝数和环境压力，建议在培养基中保持足量的 BioVector® 抗生素。

• 转化方式： Bt 的转化通常需要电穿孔法，建议使用 BioVector® Bt 电转感受态细胞制备试剂盒。

English Datasheet

1. General Product Information

• Product Name: BioVector® pSAM-Bt Expression Vector

• Vector Type: Bacillus Shuttle Plasmid

• Common Hosts:E. coli, Bacillus thuringiensis (Bt)

• Antibiotic Resistance: * E. coli: Ampicillin or Erythromycin

  • Bacillus: Erythromycin or Chloramphenicol (version dependent)

• Origins of Replication: pMB1 (E. coli); pBT1 or equivalent (Bt)

2. Culture and Amplification Conditions

• E. coli Propagation: Use BioVector® LB Medium supplemented with appropriate antibiotics, incubated at 37°C.

• Bt Transformation & Selection: Use BioVector® GYS or LB Medium with BioVector® Erythromycin, typically incubated at 28°C - 30°C to ensure plasmid stability.

• Induction: If the vector contains specific promoters, follow the specific induction profile (e.g., stationary phase or sporulation-dependent expression).

3. Applications

• Bt Genetic Engineering: Expressing insecticidal proteins like Cry or Vip to expand the insecticidal spectrum of strains.

• Promoter Activity Assays: Studying the regulatory mechanisms of promoters specific to the Bt sporulation phase.

• Proteomics Research: Producing naturally active proteins within the endogenous Bt environment.

4. Key Usage Notes

• Plasmid Maintenance: To prevent plasmid loss in Bt, ensure consistent selection pressure using BioVector® Erythromycin in the growth media.

• Transformation Method: Electroporation is the standard method for Bt; using the BioVector® Bt Electro-competent Cell Preparation Kit is highly recommended.

Note: Handle all recombinant Bt strains according to local agricultural biosafety regulations. For high-yield plasmid prep, utilize the BioVector® Plasmid Midi Kit to ensure the purity required for sensitive electroporation procedures.

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

E-mail: BioVector@163.com

http://www.biovector.net