BioVector® pBIG1 & pBIG2 biGbac 系统二级载体 / pBIG1 & pBIG2 biGbac Level 2 Vectors

通用定义 / General Definition:

BioVector® pBIG1 和 pBIG2 是 biGbac 多基因表达系统中的二级受体载体（Level 2 vectors）。在 biGbac 工作流程中，这些载体通过 Gibson Assembly（吉布森组装） 技术，接收来自多个 pLIB（一级载体）的表达盒。pBIG1 能够容纳多达 5 个基因表达盒，而 pBIG2 则用于通过进一步组装来自 pBIG1 的片段来构建包含 10 个甚至更多基因的超大型复合物。这些载体包含杆状病毒侧翼序列（$Tn7$ 转座位点），允许目的基因组装体整合到杆状病毒基因组（Bacmid）中，用于在昆虫细胞中大规模生产蛋白质。

BioVector® pBIG1 and pBIG2 are the Level 2 receptor vectors within the biGbac multi-gene expression system. In the biGbac workflow, these vectors utilize Gibson Assembly to receive multiple expression cassettes from pLIB (Level 1) vectors. pBIG1 can accommodate up to 5 gene expression cassettes, while pBIG2 is designed to construct ultra-large complexes containing 10 or more genes by assembling fragments from pBIG1. These vectors feature baculoviral flanking sequences ($Tn7$ transposition sites), facilitating the integration of the multi-gene assembly into the baculovirus genome (Bacmid) for high-level protein production in insect cells.

BioVector® pBIG1 & pBIG2 技术说明书 (Technical Datasheet)

中文版说明书

1. 产品基本信息

• 产品名称： BioVector® pBIG1 / pBIG2 (biGbac Level 2) 载体

• 系统层级： 二级组装受体载体

• 抗性基因： * pBIG1： 卡那霉素 (Kanamycin)

  • pBIG2： 壮观霉素 (Spectinomycin)

• 核心元件：$Tn7L$ & $Tn7R$ 转座末端、Gentamicin（庆大霉素，用于 Bacmid 筛选标记）

2. 培养与扩增条件

• 宿主菌： BioVector® DH5-$\alpha$ 或 PIR1/PIR2（取决于具体复制子）。

• 培养基： BioVector® LB 培养基。

• 抗生素： 按载体类型添加相应的卡那霉素 (50 ug/mL) 或壮观霉素 (100 ug/mL)。

• 环境： 37 摄氏度，恒温振荡。

3. 载体应用

• 大分子复合物表达： 构建用于冷冻电镜 (Cryo-EM) 结构研究的多亚基复合物。

• 全途径生物合成： 在昆虫细胞中重组完整的外源代谢途径。

• 疫苗研发： 表达包含多种病毒抗原的类病毒颗粒 (VLPs)。

4. 注意事项

• 组装逻辑： pBIG1 通常接收 5 个含有不同接头序列的 pLIB 插入片段。确保 pLIB 载体的编号与 pBIG1 的预设接头完全匹配。

• 转座验证： 组装完成后，需将质粒转化入 DH10Bac 感受态细胞进行蓝白斑筛选，验证基因组整合情况。

English Datasheet

1. General Product Information

• Product Name: BioVector® pBIG1 / pBIG2 (biGbac Level 2) Vectors

• System Hierarchy: Level 2 Assembly Receptor Vectors

• Antibiotic Resistance: * pBIG1: Kanamycin

  • pBIG2: Spectinomycin

• Core Elements:$Tn7L$ & $Tn7R$ transposition sites, Gentamicin resistance gene (for Bacmid selection marker).

2. Culture and Amplification Conditions

• Host Strains: BioVector® DH5-$\alpha$ or PIR1/PIR2 (depending on the specific replicon).

• Culture Medium: BioVector® LB Medium.

• Antibiotic Supplement: Kanamycin (50 ug/mL) or Spectinomycin (100 ug/mL) as per vector type.

• Incubation: 37 degrees Celsius with constant shaking.

3. Applications

• Large Complex Expression: Constructing multi-subunit complexes for structural biology studies using Cryo-EM.

• Full Pathway Biosynthesis: Reconstituting entire exogenous metabolic pathways in insect cell hosts.

• Vaccine Development: Expressing virus-like particles (VLPs) containing multiple viral antigens.

4. Key Usage Notes

• Assembly Logic: pBIG1 typically receives 5 pLIB inserts with specific flanking adapters. Ensure that the pLIB vector numbering matches the designated adapter logic within the pBIG1 target.

• Transposition Verification: Once Level 2 assembly is complete, the resulting plasmid must be transformed into DH10Bac competent cells for blue/white screening to confirm successful integration into the Bacmid.

Note: Due to the large size of the final assembled constructs (often >20 kb), BioVector® recommends using high-fidelity enzymes and optimized Gibson Assembly mixes. Always verify the integrity of the junctions via BioVector® long-read sequencing or restriction digestion mapping.

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