BioVector® pBHG-fiber5/35 腺病毒骨架载体 / BioVector® pBHG-fiber5/35 Adenoviral Backbone Plasmid

通用定义 / General Definition:

BioVector® pBHG-fiber5/35 是一种专门用于构建嵌合型腺病毒系统的骨架质粒。它基于 Ad5 腺病毒基因组，但关键性的 Fiber（纤维蛋白） 基因被改造为包含 Ad5 和 Ad35 的嵌合体（通常是 Ad5 的尾部结合 Ad35 的轴部和头部）。这种设计改变了病毒的趋向性（Tropism），使其不再依赖于常规的 CAR 受体，而是通过 CD46 受体感染细胞。这对于转染 CAR 表达较低的细胞（如造血干细胞、免疫细胞和某些肿瘤细胞）至关重要。

BioVector® pBHG-fiber5/35 is an adenoviral backbone plasmid specialized for the construction of chimeric adenovirus systems. Based on the Ad5 genome, its key Fiber gene is engineered as a chimera containing components of both Ad5 and Ad35 (typically Ad5 tail and Ad35 shaft/knob). This modification shifts the viral tropism from the standard CAR receptor to the CD46 receptor. This is essential for transducing cells with low CAR expression, such as hematopoietic stem cells, immune cells, and certain malignant cells.

BioVector® pBHG-fiber5/35 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® pBHG-fiber5/35 骨架质粒

• 病毒血清型： 嵌合型 Ad5/f35

• 受体趋向性： CD46 (人)

• 抗性基因： 氨苄青霉素 (Ampicillin) 或 结合特定体系的抗性

• 克隆方法： 与含有目的基因的 BioVector® 穿梭质粒在 HEK293 细胞或大肠杆菌内进行重组。

2. 培养与扩增条件

• 宿主菌： BioVector® Stbl3 或 DH5$\alpha$（建议使用高保真菌株以防止大质粒重组）。

• 培养基： BioVector® LB 培养基

• 抗生素添加： BioVector® 氨苄青霉素 (100 $\mu$g/mL)

• 培养环境： 30°C - 37°C 恒温振荡培养（30°C 有助于维持大质粒稳定性）。

3. 载体应用

• 难转染细胞感染： 针对 CAR 缺失的 T 细胞、B 细胞及干细胞的高效基因传递。

• 腺病毒包装： 作为骨架载体，用于产生具有 5/35 嵌合特性的复制缺陷型腺病毒。

• 肿瘤靶向研究： 利用 CD46 在多种癌细胞中高表达的特性进行靶向治疗研究。

4. 注意事项

• 质粒大小： 该载体较大，提取时需使用 BioVector® 大质粒提取试剂盒以确保完整性。

• 生物安全性： 包装出的病毒具有感染人类细胞的能力，必须在二级生物安全柜 (BSL-2) 中操作。

English Datasheet

1. General Product Information

• Product Name: BioVector® pBHG-fiber5/35 Backbone Plasmid

• Viral Serotype: Chimeric Ad5/f35

• Receptor Tropism: CD46 (Human)

• Antibiotic Resistance: Ampicillin (or specific to system)

• Cloning Method: Recombination with a BioVector® shuttle vector containing the gene of interest (GOI) in HEK293 cells or E. coli.

2. Culture and Amplification Conditions

• Host Strains: BioVector® Stbl3 or DH5$\alpha$ (High-fidelity strains recommended to prevent large plasmid rearrangement).

• Culture Medium: BioVector® LB Medium

• Antibiotic Supplement: BioVector® Ampicillin (100 $\mu$g/mL)

• Incubation: 30°C - 37°C constant shaking (30°C is preferred for maintaining stability of large constructs).

3. Applications

• Refractory Cell Transduction: High-efficiency gene delivery to CAR-deficient cells such as T-cells, B-cells, and stem cells.

• Adenovirus Packaging: Serves as the backbone for generating replication-deficient adenoviruses with 5/35 chimeric properties.

• Cancer Targeting: Exploiting the high expression of CD46 in various malignancies for targeted therapy research.

4. Key Usage Notes

• Plasmid Size: Due to its large size, use a BioVector® Large Plasmid Extraction Kit to ensure structural integrity.

• Biosafety: The resulting packaged virus is capable of infecting human cells; all downstream viral work must be conducted under Biosafety Level 2 (BSL-2) protocols.

Note: For maximum viral titer, ensure the purity of BioVector® pBHG-fiber5/35 before transfection. Use BioVector® Competent Cells for stable propagation of this complex backbone.

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