pDS05 BioVector®温度敏感型自杀型载体 (Suicide Vector) / 等位基因交换载体

BioVector®  pDS05自杀型载体 (Suicide Vector) / 等位基因交换载体

• 质粒名称: pDS05

• 质粒类型: 自杀型载体 (Suicide Vector) / 等位基因交换载体。

• 用途: 用于细菌基因组的无痕缺失 (Scarless Deletion)、点突变或基因敲入。

• 衍生来源: 基于 pWM91 或类似的 R6K 复制子载体改造。 [x]

• 复制子: R6Kγ ori。该复制子依赖于 pir 蛋白。在缺乏 pir 基因的宿主中无法复制，从而迫使质粒整合进染色体。 [x]

• 反向筛选标记: sacB。在含蔗糖的培养基上，该基因产物对细菌有毒，用于筛选发生第二次交叉（二桥）并丢失质粒骨架的克隆。 [x]

• 抗性标记: 氨苄青霉素 (Ampicillin) 或其他（取决于具体变体，最常用为 AmpR）。

• 转移原点: oriT。允许通过接合 (Conjugation) 从供体菌（如 S17-1 λpir）转移到受体菌中。 [x]

• 无痕编辑: 实验结束后，染色体上不会留下任何抗性标记或疤痕序列。

• 宿主范围: 广泛用于革兰氏阴性菌，特别是 弧菌属 (Vibrio) 和 肠道杆菌。

Plasmid Datasheet - English Version

• Plasmid Name: pDS05

• Plasmid Type:Suicide Vector / Allelic Exchange Vector.

• Purpose: Used for scarless gene deletion, site-directed mutagenesis, or gene knock-ins in bacterial genomes.

• Origin: Often a derivative of the pWM91 lineage or other R6K-based suicide plasmids. [x]

• Replicon:R6Kγ ori. Requires the pir gene product for replication. It acts as a "suicide" plasmid in standard laboratory strains, forcing integration into the host chromosome. [x]

• Counter-selection Marker:sacB. Encodes levansucrase; confers sensitivity to sucrose. This allows for the selection of the second crossover event where the plasmid backbone is excised. [x]

• Selectable Marker: Usually Ampicillin resistance (AmpR).

• Mobilization:oriT (Origin of Transfer). Enables mobilization via conjugation from helper strains like E. coli S17-1 λpir into the target recipient. [x]

• Scarless Modification: Ensures that no antibiotic resistance markers remain in the genome after the successful completion of the two-step recombination.

• Target Species: Commonly utilized in studies of Vibrio cholerae, Yersinia pestis, and other Enterobacteriaceae.

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BioVector NTCC Inc.

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