BioVector? pENTR11Dual 入門克隆載體 / BioVector? pENTR11Dual Entry Vector

通用定義 / General Definition:

BioVector? pENTR11Dual 是一種高度靈活的 Gateway? 入門載體，旨在通過 $attL1$ 和 $attL2$ 重組位點高效構(gòu)建供體克隆。其“雙重（Dual）”特性通常體現(xiàn)在其多克隆位點（MCS）的設(shè)計上，允許使用傳統(tǒng)的限制性內(nèi)切酶克隆方法插入目的基因，同時其兩側(cè)的 Gateway 位點支持后續(xù)通過 LR 反應將基因快速轉(zhuǎn)移至各類表達載體（Destination Vectors）。它常用于高通量蛋白表達、多基因共表達以及跨物種的基因功能研究。

BioVector? pENTR11Dual is a highly versatile Gateway? Entry Vector designed for the efficient construction of donor clones via $attL1$ and $attL2$ recombination sites. Its "Dual" characteristic typically refers to its multiple cloning site (MCS) design, which allows for the insertion of target genes using traditional restriction enzyme-based cloning, while the flanking Gateway sites facilitate the rapid transfer of the gene into various Destination Vectors through LR reactions. It is widely used in high-throughput protein expression, multi-gene co-expression, and cross-species functional genomics.

BioVector? pENTR11Dual 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? pENTR11Dual 入門載體

• 克隆方法： 限制性內(nèi)切酶克隆 或 Gateway? 重組技術(shù)

• 載體類型： 入門載體 (Entry Vector)

• 抗性基因： 卡那霉素 (Kanamycin)

• 篩選標記：$ccdB$ 致死基因（用于陰性篩選）

2. 培養(yǎng)與擴增條件

• 宿主菌： BioVector? cccB 敏感型菌株（如 DH5$\alpha$）用于含有目的基因的克??；BioVector? $ccdB$ 抗性菌株（如 $ccdB$ Survival?）用于空載體擴增。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基

• 抗生素添加： BioVector? 卡那霉素 (50 $\mu$g/mL)

• 培養(yǎng)環(huán)境： 37°C，恒溫振蕩培養(yǎng)

3. 載體應用

• 基因穿梭： 將目的基因從 BioVector? pENTR11Dual 轉(zhuǎn)移至各種含有 $attR$ 位點的表達載體中。

• 雙啟動子系統(tǒng)： 支持在不同的宿主系統(tǒng)中通過重組實現(xiàn)高效表達。

• 高通量克?。?/strong> 適用于大規(guī)模功能基因組篩選。

4. 注意事項

• 負向篩選： 載體含有 $ccdB$ 基因，常規(guī) $E. coli$ 轉(zhuǎn)化前請確保目的基因已成功替換該致死基因。

• 序列驗證： 建議使用 BioVector? M13 Forward 和 M13 Reverse 引物對插入片段進行測序驗證。

English Datasheet

1. General Product Information

• Product Name: BioVector? pENTR11Dual Entry Vector

• Cloning Method: Restriction Enzyme Cloning or Gateway? Recombination Technology

• Vector Type: Entry Vector

• Antibiotic Resistance: Kanamycin

• Selection Marker:$ccdB$ lethal gene (for negative selection)

2. Culture and Amplification Conditions

• Host Strains: BioVector? cccB-sensitive strains (e.g., DH5$\alpha$) for recombinants; BioVector? $ccdB$ Survival? strains for empty vector propagation.

• Culture Medium: BioVector? LB Medium

• Antibiotic Supplement: BioVector? Kanamycin (50 $\mu$g/mL)

• Incubation: 37°C, constant shaking

3. Applications

• Gene Shuttling: Transferring target genes from BioVector? pENTR11Dual into various Destination Vectors containing $attR$ sites.

• Dual Promoter Systems: Facilitating high-efficiency expression across different host systems via recombination.

• High-Throughput Cloning: Ideal for large-scale functional genomics screening.

4. Key Usage Notes

• Negative Selection: The vector contains the $ccdB$ gene; ensure the target gene has successfully replaced this lethal cassette before transformation into standard $E. coli$ competent cells.

• Sequence Verification: It is recommended to verify the insert using BioVector? M13 Forward and M13 Reverse sequencing primers.

Note: For optimal cloning efficiency and plasmid yield, use BioVector? LB Medium and BioVector? competent cells. Always perform restriction enzyme digestion or sequencing to confirm the orientation and integrity of your insert within the BioVector? pENTR11Dual backbone.

Supplier生產(chǎn)廠家: BioVector NTCC典型培養(yǎng)物保藏中心

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