BioVector? NTCC? pRNAT-CMV3.1-Puro真核短發(fā)夾RNA表達(dá)載體BioVector?

pRNAT-CMV3.1-Puro BioVector? plasmid

基本信息

質(zhì)粒名稱： pRNAT-CMV3.1-Puro

載體類型： 真核表達(dá)載體

主要用途：

在哺乳動(dòng)物細(xì)胞中驅(qū)動(dòng) shRNA 或其他短 RNA 的表達(dá)

在細(xì)胞系中進(jìn)行穩(wěn)定表達(dá)篩選（通過嘌呤霉素抗性基因）

骨架來源： pRNAT 系列載體

關(guān)鍵特征（非操作性概述）

啟動(dòng)子：CMV（增強(qiáng)型巨細(xì)胞病毒啟動(dòng)子）
用于在多種哺乳動(dòng)物細(xì)胞中實(shí)現(xiàn)高水平表達(dá)。

shRNA 表達(dá)框架：
設(shè)計(jì)用于驅(qū)動(dòng)人工合成的短發(fā)夾 RNA（shRNA）表達(dá)；通常包含 RNA polymerase III 啟動(dòng)子（如 U6 或 H1），具體視構(gòu)建版本而定。

篩選標(biāo)記：嘌呤霉素抗性基因（PuroR）
允許在真核細(xì)胞中進(jìn)行穩(wěn)定轉(zhuǎn)染篩選。

報(bào)告基因（部分構(gòu)建具有）：
某些版本包含熒光蛋白（例如 GFP）以便觀察轉(zhuǎn)染效率（依供應(yīng)商不同）。

復(fù)制與選擇標(biāo)記（細(xì)菌側(cè)）

大腸桿菌復(fù)制起點(diǎn)： 常見 ColE1 類型

細(xì)菌抗性基因： 氨芐青霉素抗性基因（AmpR）

(注意：此處僅為特性描述，不包含任何實(shí)驗(yàn)步驟、培養(yǎng)條件或操作指南。)

質(zhì)粒大小

一般約 5–7 kb（具體視版本或是否插入 RNA 序列而變化）

常見應(yīng)用（高層級(jí)說明）

RNA 干擾（RNAi）研究

基因功能驗(yàn)證

通過嘌呤霉素篩選建立穩(wěn)定細(xì)胞系

shRNA 媒介的基因沉默機(jī)制研究

General Information

Plasmid Name: pRNAT-CMV3.1-Puro

Type: Mammalian expression vector designed for RNA interference (shRNA expression)

Backbone: pRNAT-CMV3.1 series

Size: ~6–7 kb (varies by insert; check vendor’s specific map)

Key Features (High-Level)

Promoter for shRNA cassette: CMV promoter

Strong, constitutive promoter suitable for many mammalian cell types

Fluorescent Marker: Often contains EGFP or similar reporter for tracking transfection efficiency (depends on the version)

Selection Marker (Mammalian): Puromycin resistance gene

Enables selection of stably transfected mammalian cells

Selection Marker (Bacterial): Typically Ampicillin resistance (varies by lot/source)

Intended Applications

Expression of short hairpin RNA (shRNA) in mammalian cells

Gene knockdown studies

Generation of stable RNAi cell lines

Visualization of transfection efficiency via fluorescent reporter

Expression Elements (Overview)

CMV promoter → shRNA expression cassette

Pol III promoter (e.g., U6) in some versions for RNA polymerase III-driven shRNA transcription

Polyadenylation signal for proper transcript termination (CMV-driven sequences)

Map Components (Typical, High-Level)

CMV promoter

Multiple cloning site (MCS) for shRNA or hairpin insertion

Puromycin resistance gene

EGFP reporter (if included in that version)

Origin of replication (bacterial)

Ampicillin resistance gene (common but may vary)

Biosafety Classification

Generally BSL-1, as a non-replicating plasmid

Final biosafety depends on inserted gene, host cell line, and institutional rules

Storage & Stability (Non-procedural)

Stable when stored under manufacturer-recommended long-term conditions

Avoid repeated freeze–thaw cycles (general plasmid guidance)

Quality Control (Vendor-Level)

Sequence verification of backbone

Antibiotic resistance confirmation

Host strain identity verification

Purity and sterility testing depending on supplier

Supplier來源:BioVector NTCC Inc.
Email: [email protected]
Website網(wǎng)址: http://www.nedfriskphoto.com