BioVector? pLenti-CMV-IRS4-Myc/His-Puro 慢病毒表達質(zhì)粒 / BioVector? pLenti-CMV-IRS4-Myc/His-Puro Lentiviral Expression Plasmid

通用定義 / General Definition:BioVector? pLenti-CMV-IRS4-Myc/His-Puro 是一種基于第三代慢病毒系統(tǒng)的哺乳動物表達載體，專門設(shè)計用于在靶細胞中實現(xiàn) IRS4 (胰島素受體底物 4) 蛋白的高效整合與穩(wěn)定表達。該質(zhì)粒利用強力的 CMV 啟動子 驅(qū)動 IRS4 的轉(zhuǎn)錄，并在其 C 端融合了 Myc 和 His 雙標簽，方便通過免疫印跡 (WB)、免疫共沉淀 (Co-IP) 或親和層析進行蛋白檢測與純化。載體同時攜帶 Puro (嘌呤霉素) 抗性基因，允許研究人員篩選建立穩(wěn)定表達 IRS4 的細胞株。IRS4 是胰島素和細胞因子信號轉(zhuǎn)導中的重要接頭蛋白，常在多種癌癥和代謝研究中被深入探討。

BioVector? pLenti-CMV-IRS4-Myc/His-Puro is a third-generation lentiviral expression vector designed for the stable integration and high-level expression of IRS4 (Insulin Receptor Substrate 4) in mammalian cells. The vector utilizes a robust CMV promoter to drive IRS4 transcription and features C-terminal Myc and His tags to facilitate protein detection, pull-down assays, or purification. Additionally, it contains a Puro (Puromycin) resistance marker for the selection of stable cell lines. IRS4 acts as a critical adaptor protein in insulin and cytokine signaling pathways and is frequently investigated in oncogenesis and metabolic disorder research.

BioVector? pLenti-CMV-IRS4-Myc/His-Puro 技術(shù)說明書 (Technical Datasheet)

中文版說明書 (Chinese Datasheet)

1. 質(zhì)?；拘畔?/strong>

• 載體類型： 慢病毒表達載體 (Lentiviral Expression Vector)

• 啟動子： CMV (Cytomegalovirus) 啟動子

• 插入片段： 人或小鼠 IRS4 (Insulin Receptor Substrate 4)

• 融合標簽： C-terminal Myc-His 雙標簽

• 篩選標記： Puromycin (嘌呤霉素)

• 克隆位點： 通常位于 MCS 多克隆位點

• 細菌抗性： 氨芐青霉素 (Ampicillin, 100 mg/L)

2. 核心生物學功能

• 穩(wěn)定轉(zhuǎn)導： 慢病毒系統(tǒng)可高效感染分裂和非分裂細胞，并將 IRS4 基因組整合至宿主染色體。

• 信號通路研究： 探索 IRS4 對 PI3K/Akt 和 MAPK 信號通路的調(diào)節(jié)作用。

• 蛋白互作： 利用 Myc/His 標簽通過質(zhì)譜或 Co-IP 鑒定 IRS4 的下游結(jié)合伙伴。

3. 實驗應(yīng)用方案

• 病毒包裝： 需配合包裝系統(tǒng)（如 pMD2.G 和 psPAX2）在 293T 細胞中進行病毒生產(chǎn)。

• 穩(wěn)轉(zhuǎn)株建立： 感染靶細胞后，通過梯度濃度的嘌呤霉素進行抗性篩選。

• 功能驗證： 通過 Western Blot 驗證 IRS4-Myc/His 的表達情況。

4. 注意事項

• 生物安全： 包裝后的病毒顆粒必須在生物安全二級 (BSL-2) 環(huán)境下操作。

• 片段長度： 由于 IRS4 編碼區(qū)較長，請確保病毒包裝效率，必要時進行病毒濃縮。

English Datasheet

1. Basic Plasmid Information

• Vector Type: Lentiviral Expression Vector

• Promoter: CMV (Cytomegalovirus)

• Insert: Human or Mouse IRS4 (Insulin Receptor Substrate 4)

• Fusion Tags: C-terminal Myc-His dual tags

• Selection Marker: Puromycin (Puro)

• Bacterial Selection: Ampicillin (100 mg/L)

• Replicon: pUC ori

2. Key Biological Functions

• Stable Expression: Capable of mediating long-term stable expression in a wide range of mammalian cell types via genomic integration.

• Signaling Analysis: Serving as a tool to investigate the impact of IRS4 over-expression on insulin signaling and cell proliferation.

• Tag Utility: The Myc/His tandem tags enable high-specificity detection using anti-Myc/His antibodies and simplified protein purification via IMAC (Immobilized Metal Affinity Chromatography).

3. Applications

• Lentiviral Production: Standard transfection into 293T cells using a 3-plasmid packaging system.

• Stable Line Generation: Selection of transduced cells using Puromycin following viral infection.

• Pathway Profiling: Analyzing phosphorylation patterns of Akt or ERK in response to IRS4 expression.

4. Key Usage Notes

• Biosafety: Handle as BSL-2 material once packaged into viral particles.

• Sequence Integrity: BioVector? recommends verifying the IRS4-Myc-His junction via Sanger sequencing to ensure the reading frame is correct.

注意 / Note: 質(zhì)粒應(yīng)儲存在 -20 攝氏度。在使用 CMV 啟動子時，需注意某些特定的細胞系（如干細胞）可能會發(fā)生啟動子沉默現(xiàn)象。Store at -20°C. When using the CMV promoter, be aware that promoter silencing can occur in certain cell types, such as stem cells or primary neurons.

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