BioVector® p423 GAL1 酵母表达载体 / BioVector® p423 GAL1 Yeast Expression Vector

通用定义 / General Definition:

BioVector® p423 GAL1 是一种用于酿酒酵母（Saccharomyces cerevisiae）的高拷贝、诱导型表达载体。该载体属于 pRS42x 系列，包含 $2\mu$ 起始位点（用于高拷贝维持）和 $HIS3$ 营养缺陷型筛选标记。其核心特征是采用了 $GAL1$ 启动子，这使得目的基因的表达受半乳糖诱导并受葡萄糖严谨阻遏。它广泛应用于酵母中的蛋白超表达、功能基因组学以及毒性蛋白的受控表达研究。

BioVector® p423 GAL1 is a high-copy, inducible expression vector designed for use in Saccharomyces cerevisiae. Part of the pRS42x series, it features the $2\mu$ origin (for high-copy maintenance) and the $HIS3$ auxotrophic marker for selection. The core component is the $GAL1$ promoter, which allows for stringent control of gene expression—induced by galactose and repressed by glucose. It is widely used for protein overexpression, functional genomics, and the controlled expression of toxic proteins in yeast.

BioVector® p423 GAL1 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® p423 GAL1 表达载体

• 载体类型： 酵母高拷贝表达载体 (High-copy episomal vector)

• 启动子： BioVector® $GAL1$ 诱导型启动子

• 筛选标记： * 大肠杆菌： 氨苄青霉素 (Ampicillin)

  • 酵母：$HIS3$ (组氨酸营养缺陷型)

• 复制子：$2\mu$ (酵母); $pMB1$ (大肠杆菌)

2. 培养与扩增条件

• 大肠杆菌扩增： 使用 BioVector® LB 培养基，添加 100 $\mu$g/mL BioVector® 氨苄青霉素，37°C 培养。

• 酵母筛选： 使用 BioVector® 组氨酸缺陷型合成完全培养基 (SC-His)。

• 表达诱导： 先在含葡萄糖的 BioVector® SC-His 中扩增，待细胞达到对数生长期后，转入含 2% 半乳糖的 BioVector® 诱导培养基。

3. 载体应用

• 代谢调控： 通过半乳糖诱导实现目的蛋白的大量表达。

• 毒性蛋白研究： 利用 $GAL1$ 启动子在葡萄糖中的高度抑制特性，克隆对酵母有毒的基因。

• 多质粒共转化： 可与其他带有不同标记（如 $URA3, LEU2, TRP1$）的 BioVector® p42x 系列载体配合使用。

4. 注意事项

• 葡萄糖抑制： 即使是微量的葡萄糖也会显著抑制 $GAL1$ 启动子的活性。诱导前建议彻底清洗细胞。

• 拷贝数：$2\mu$ 载体在细胞内通常维持在 10-40 个拷贝，可能导致表达水平波动。

English Datasheet

1. General Product Information

• Product Name: BioVector® p423 GAL1 Expression Vector

• Vector Type: Yeast High-copy Episomal Vector

• Promoter: BioVector® $GAL1$ Inducible Promoter

• Selection Markers: * E. coli: Ampicillin

  • Yeast:$HIS3$ (Histidine auxotrophy)

• Origins of Replication:$2\mu$ (Yeast); $pMB1$ (E. coli)

2. Culture and Amplification Conditions

• E. coli Propagation: Use BioVector® LB Medium supplemented with 100 $\mu$g/mL BioVector® Ampicillin, incubated at 37°C.

• Yeast Selection: Use BioVector® Synthetic Complete Medium lacking Histidine (SC-His).

• Induction Profile: Pre-culture in BioVector® SC-His with Glucose; shift to BioVector® induction medium containing 2% Galactose during the log phase.

3. Applications

• Metabolic Control: Large-scale protein expression via galactose induction.

• Toxic Protein Expression: Utilizing the tight repression of the $GAL1$ promoter in glucose to clone genes detrimental to yeast growth.

• Co-transformation: Compatible with other BioVector® p42x series vectors carrying different markers (e.g., $URA3, LEU2, TRP1$).

4. Key Usage Notes

• Glucose Repression: Even trace amounts of glucose can significantly inhibit $GAL1$ promoter activity. Ensure cells are thoroughly washed before switching to induction medium.

• Copy Number: As a $2\mu$ based vector, it typically maintains 10-40 copies per cell, which may lead to variable expression levels across a population.

Note: For optimal transformation efficiency, use BioVector® Yeast Competent Cell Pre-kits. To maintain the integrity of the GAL induction system, consistently use BioVector® SC-His Medium and BioVector® Ampicillin for plasmid maintenance.

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