BioVector? p423 GAL1 酵母表達(dá)載體 / BioVector? p423 GAL1 Yeast Expression Vector

通用定義 / General Definition:

BioVector? p423 GAL1 是一種用于釀酒酵母（Saccharomyces cerevisiae）的高拷貝、誘導(dǎo)型表達(dá)載體。該載體屬于 pRS42x 系列，包含 $2\mu$ 起始位點(diǎn)（用于高拷貝維持）和 $HIS3$ 營(yíng)養(yǎng)缺陷型篩選標(biāo)記。其核心特征是采用了 $GAL1$ 啟動(dòng)子，這使得目的基因的表達(dá)受半乳糖誘導(dǎo)并受葡萄糖嚴(yán)謹(jǐn)阻遏。它廣泛應(yīng)用于酵母中的蛋白超表達(dá)、功能基因組學(xué)以及毒性蛋白的受控表達(dá)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 載體技術(shù)說(shuō)明書 (Plasmid Datasheet)

中文版說(shuō)明書

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? p423 GAL1 表達(dá)載體

• 載體類型： 酵母高拷貝表達(dá)載體 (High-copy episomal vector)

• 啟動(dòng)子： BioVector? $GAL1$ 誘導(dǎo)型啟動(dòng)子

• 篩選標(biāo)記： * 大腸桿菌： 氨芐青霉素 (Ampicillin)

  • 酵母：$HIS3$ (組氨酸營(yíng)養(yǎng)缺陷型)

• 復(fù)制子：$2\mu$ (酵母); $pMB1$ (大腸桿菌)

2. 培養(yǎng)與擴(kuò)增條件

• 大腸桿菌擴(kuò)增： 使用 BioVector? LB 培養(yǎng)基，添加 100 $\mu$g/mL BioVector? 氨芐青霉素，37°C 培養(yǎng)。

• 酵母篩選： 使用 BioVector? 組氨酸缺陷型合成完全培養(yǎng)基 (SC-His)。

• 表達(dá)誘導(dǎo)： 先在含葡萄糖的 BioVector? SC-His 中擴(kuò)增，待細(xì)胞達(dá)到對(duì)數(shù)生長(zhǎng)期后，轉(zhuǎn)入含 2% 半乳糖的 BioVector? 誘導(dǎo)培養(yǎng)基。

3. 載體應(yīng)用

• 代謝調(diào)控： 通過(guò)半乳糖誘導(dǎo)實(shí)現(xiàn)目的蛋白的大量表達(dá)。

• 毒性蛋白研究： 利用 $GAL1$ 啟動(dòng)子在葡萄糖中的高度抑制特性，克隆對(duì)酵母有毒的基因。

• 多質(zhì)粒共轉(zhuǎn)化： 可與其他帶有不同標(biāo)記（如 $URA3, LEU2, TRP1$）的 BioVector? p42x 系列載體配合使用。

4. 注意事項(xiàng)

• 葡萄糖抑制： 即使是微量的葡萄糖也會(huì)顯著抑制 $GAL1$ 啟動(dòng)子的活性。誘導(dǎo)前建議徹底清洗細(xì)胞。

• 拷貝數(shù)：$2\mu$ 載體在細(xì)胞內(nèi)通常維持在 10-40 個(gè)拷貝，可能導(dǎo)致表達(dá)水平波動(dòng)。

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.

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