BioVector? pBIG1 & pBIG2 biGbac 系統(tǒng)二級(jí)載體 / pBIG1 & pBIG2 biGbac Level 2 Vectors

通用定義 / General Definition:

BioVector? pBIG1 和 pBIG2 是 biGbac 多基因表達(dá)系統(tǒng)中的二級(jí)受體載體（Level 2 vectors）。在 biGbac 工作流程中，這些載體通過 Gibson Assembly（吉布森組裝） 技術(shù)，接收來自多個(gè) pLIB（一級(jí)載體）的表達(dá)盒。pBIG1 能夠容納多達(dá) 5 個(gè)基因表達(dá)盒，而 pBIG2 則用于通過進(jìn)一步組裝來自 pBIG1 的片段來構(gòu)建包含 10 個(gè)甚至更多基因的超大型復(fù)合物。這些載體包含桿狀病毒側(cè)翼序列（$Tn7$ 轉(zhuǎn)座位點(diǎn)），允許目的基因組裝體整合到桿狀病毒基因組（Bacmid）中，用于在昆蟲細(xì)胞中大規(guī)模生產(chǎn)蛋白質(zhì)。

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 技術(shù)說明書 (Technical Datasheet)

中文版說明書

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? pBIG1 / pBIG2 (biGbac Level 2) 載體

• 系統(tǒng)層級(jí)： 二級(jí)組裝受體載體

• 抗性基因： * pBIG1： 卡那霉素 (Kanamycin)

  • pBIG2： 壯觀霉素 (Spectinomycin)

• 核心元件：$Tn7L$ & $Tn7R$ 轉(zhuǎn)座末端、Gentamicin（慶大霉素，用于 Bacmid 篩選標(biāo)記）

2. 培養(yǎng)與擴(kuò)增條件

• 宿主菌： BioVector? DH5-$\alpha$ 或 PIR1/PIR2（取決于具體復(fù)制子）。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基。

• 抗生素： 按載體類型添加相應(yīng)的卡那霉素 (50 ug/mL) 或壯觀霉素 (100 ug/mL)。

• 環(huán)境： 37 攝氏度，恒溫振蕩。

3. 載體應(yīng)用

• 大分子復(fù)合物表達(dá)： 構(gòu)建用于冷凍電鏡 (Cryo-EM) 結(jié)構(gòu)研究的多亞基復(fù)合物。

• 全途徑生物合成： 在昆蟲細(xì)胞中重組完整的外源代謝途徑。

• 疫苗研發(fā)： 表達(dá)包含多種病毒抗原的類病毒顆粒 (VLPs)。

4. 注意事項(xiàng)

• 組裝邏輯： pBIG1 通常接收 5 個(gè)含有不同接頭序列的 pLIB 插入片段。確保 pLIB 載體的編號(hào)與 pBIG1 的預(yù)設(shè)接頭完全匹配。

• 轉(zhuǎn)座驗(yàn)證： 組裝完成后，需將質(zhì)粒轉(zhuǎn)化入 DH10Bac 感受態(tài)細(xì)胞進(jìn)行藍(lán)白斑篩選，驗(yàn)證基因組整合情況。

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.

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