BioVector? pLIB biGbac 系統(tǒng)載體 / BioVector? pLIB biGbac System Vector

通用定義 / General Definition:BioVector? pLIB 在 biGbac 系統(tǒng)中扮演著核心角色。biGbac 是一種基于 Gibson Assembly（吉布森組裝） 的高效多基因表達(dá)平臺(tái)，專門用于在昆蟲細(xì)胞（如 Sf9, High Five）中通過(guò)桿狀病毒表達(dá)系統(tǒng)表達(dá)復(fù)雜的大型多亞基蛋白質(zhì)復(fù)合物。在 biGbac 工作流程中，pLIB 載體作為一級(jí)克隆載體（Level 1），用于接收和儲(chǔ)存單個(gè)基因。隨后，多個(gè) pLIB 載體中的基因片段可以被組裝進(jìn)二級(jí)載體（Level 2，如 pBIG1 或 pBIG2），最終實(shí)現(xiàn)多個(gè)基因的同時(shí)高水平表達(dá)。

In the biGbac system, BioVector? pLIB serves as the fundamental Level 1 cloning vector. The biGbac system is a high-efficiency multi-gene expression platform based on Gibson Assembly, specifically designed for expressing large, multi-subunit protein complexes in insect cells (e.g., Sf9, High Five) using the Baculovirus Expression Vector System (BEVS). Within the biGbac workflow, pLIB vectors hold individual genes. These gene cassettes are then assembled into Level 2 vectors (such as pBIG1 or pBIG2) to achieve the simultaneous, high-level expression of multiple genes from a single baculoviral genome.

BioVector? pLIB biGbac 技術(shù)說(shuō)明書 (Plasmid Datasheet)

中文版說(shuō)明書

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? pLIB (biGbac Level 1) 載體

• 系統(tǒng)用途： 多基因復(fù)合體表達(dá) (biGbac 系統(tǒng))

• 啟動(dòng)子： Polh (多角體蛋白啟動(dòng)子)

• 抗性基因： 氨芐青霉素 (Ampicillin)

• 克隆位點(diǎn)： 專門設(shè)計(jì)的用于 Gibson 組裝的側(cè)翼序列

2. 培養(yǎng)與擴(kuò)增條件

• 宿主菌： BioVector? DH5-alpha。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基。

• 抗生素： BioVector? 氨芐青霉素 (100 ug/mL)。

• 環(huán)境： 37 攝氏度，恒溫振蕩。

3. 載體應(yīng)用

• 多亞基蛋白組裝： 用于構(gòu)建包含 5 個(gè)、10 個(gè)甚至更多亞基的蛋白質(zhì)復(fù)合物。

• 代謝通路重建： 在昆蟲細(xì)胞中重組復(fù)雜的代謝路徑。

• 結(jié)構(gòu)生物學(xué)： 為冷凍電鏡 (Cryo-EM) 或 X 射線晶體學(xué)提供高質(zhì)量的復(fù)合體蛋白。

4. 注意事項(xiàng)

• 側(cè)翼序列： 每個(gè) pLIB 版本（如 pLIB-1, pLIB-2 等）具有特定的接頭序列，必須按照 biGbac 組裝邏輯選擇正確的載體順序。

• 序列驗(yàn)證： 在進(jìn)行二級(jí)組裝前，必須通過(guò) BioVector? 測(cè)序服務(wù)確保 pLIB 內(nèi)部的目的基因序列完全準(zhǔn)確。

English Datasheet

1. General Product Information

• Product Name: BioVector? pLIB (biGbac Level 1) Vector

• System Purpose: Multi-protein complex expression (biGbac platform)

• Promoter: Polh (Polyhedrin promoter)

• Antibiotic Resistance: Ampicillin

• Cloning Design: Flanked by specific sequences optimized for Gibson Assembly

2. Culture and Amplification Conditions

• Host Strains: BioVector? DH5-alpha.

• Culture Medium: BioVector? LB Medium.

• Antibiotic Supplement: BioVector? Ampicillin (100 ug/mL).

• Incubation: 37 degrees Celsius with constant shaking.

3. Applications

• Multi-subunit Assembly: Constructing protein complexes containing 5, 10, or more individual subunits.

• Metabolic Engineering: Reconstituting complex metabolic pathways within insect cell hosts.

• Structural Biology: Producing high-quality protein complexes for Cryo-EM or X-ray crystallography.

4. Key Usage Notes

• Flanking Sequences: Each pLIB variant (e.g., pLIB-1, pLIB-2) contains unique linker sequences. It is critical to select the correct pLIB position according to the biGbac assembly logic.

• Sequence Integrity: Before proceeding to Level 2 assembly, ensure the gene insert within pLIB is verified using BioVector? sequencing services to confirm 100% accuracy.

Note: For optimal Gibson Assembly efficiency, use the BioVector? biGbac Assembly Master Mix. When purifying pLIB plasmids for large-scale assembly, ensure high DNA concentration and purity using BioVector? Plasmid Midi Kits.

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

E-mail: [email protected]

http://www.nedfriskphoto.com