BioVector? Ki67 過表達(dá)質(zhì)粒 / BioVector? Ki67 Overexpression Plasmid

通用定義 / General Definition:

BioVector? Ki67 過表達(dá)質(zhì)粒是一種專門設(shè)計(jì)的真核表達(dá)載體，用于在哺乳動(dòng)物細(xì)胞中高效表達(dá) Ki67（由 MKI67 基因編碼） 蛋白。Ki67 是一種關(guān)鍵的增殖標(biāo)記蛋白，僅在細(xì)胞周期的活躍階段（G1、S、G2 和 M 期）表達(dá)，而在靜止期（G0 期）缺失。該過表達(dá)模型廣泛用于研究細(xì)胞增殖調(diào)控、腫瘤進(jìn)展機(jī)制、細(xì)胞周期動(dòng)力學(xué)以及作為癌癥預(yù)后研究的陽性對(duì)照。

BioVector? Ki67 Overexpression Plasmid is a specially designed eukaryotic expression vector intended for the high-level expression of the Ki67 (encoded by the MKI67 gene) protein in mammalian cells. Ki67 is a critical proliferation marker expressed exclusively during active phases of the cell cycle (G1, S, G2, and M phases) and is absent in resting cells (G0 phase). This overexpression model is widely utilized to study cell proliferation regulation, tumor progression mechanisms, cell cycle dynamics, and as a positive control in cancer prognosis research.

BioVector? Ki67 過表達(dá)質(zhì)粒技術(shù)說明書 (Plasmid Datasheet)

中文版說明書

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? Ki67 過表達(dá)質(zhì)粒

• 目的基因： 人/小鼠 MKI67 (Ki67)

• 啟動(dòng)子： BioVector? CMV 或 EF1$\alpha$ 強(qiáng)啟動(dòng)子

• 載體骨架： BioVector? pCDNA3.1(+) 或類似哺乳動(dòng)物表達(dá)骨架

• 抗性基因： * 大腸桿菌： 氨芐青霉素 (Ampicillin) 或 卡那霉素 (Kanamycin)

  • 真核篩選： 新霉素 (G418) 或 嘌呤霉素 (Puromycin)

• 標(biāo)簽： 可選 Flag, Myc, 或 GFP 標(biāo)簽（用于蛋白定位與檢測(cè)）

2. 培養(yǎng)與擴(kuò)增條件

• 宿主菌： BioVector? DH5$\alpha$ 或 Stbl3（針對(duì) Ki67 這種大片段基因建議使用高穩(wěn)定性菌株）。

• 培養(yǎng)基： BioVector? LB 培養(yǎng)基

• 抗生素添加： 按載體要求添加 BioVector? 氨芐青霉素 (100 $\mu$g/mL)。

• 培養(yǎng)環(huán)境： 37°C 恒溫振蕩培養(yǎng)。

3. 載體應(yīng)用

• 細(xì)胞增殖研究： 誘導(dǎo)靜止期細(xì)胞進(jìn)入細(xì)胞周期。

• 腫瘤模型構(gòu)建： 研究 Ki67 高表達(dá)對(duì)腫瘤生長和侵襲的影響。

• 抗體檢測(cè)驗(yàn)證： 作為 Ki67 免疫組化 (IHC) 或流式細(xì)胞術(shù) (FACS) 檢測(cè)的陽性對(duì)照。

4. 注意事項(xiàng)

• 片段大?。?/strong> Ki67 編碼區(qū)序列非常大（人源約 9-10 kb），質(zhì)?？傞L可能超過 15 kb，轉(zhuǎn)染時(shí)建議使用 BioVector? 高效大片段轉(zhuǎn)染試劑。

• 序列穩(wěn)定性： 大片段 DNA 在克隆過程中易發(fā)生重組，建議擴(kuò)增后進(jìn)行雙酶切鑒定。

English Datasheet

1. General Product Information

• Product Name: BioVector? Ki67 Overexpression Plasmid

• Target Gene: Human / Mouse MKI67 (Ki67)

• Promoter: BioVector? CMV or EF1$\alpha$ constitutive promoter

• Vector Backbone: BioVector? pCDNA3.1(+) or equivalent mammalian expression backbone

• Selection Markers: * E. coli: Ampicillin or Kanamycin

  • Eukaryotic: Neomycin (G418) or Puromycin

• Tag Options: Available with Flag, Myc, or GFP tags for protein localization and detection.

2. Culture and Amplification Conditions

• Host Strains: BioVector? DH5$\alpha$ or Stbl3 (Stbl3 is recommended for high stability of the large Ki67 insert).

• Culture Medium: BioVector? LB Medium

• Antibiotic Supplement: BioVector? Ampicillin (100 $\mu$g/mL) as per backbone requirements.

• Incubation: 37°C with constant shaking.

3. Applications

• Proliferation Research: Inducing quiescent cells to re-enter the cell cycle.

• Tumorigenesis Models: Investigating the impact of Ki67 upregulation on tumor growth and invasiveness.

• Assay Validation: Serving as a positive control for Ki67 Immunohistochemistry (IHC) or Flow Cytometry (FACS) detection.

4. Key Usage Notes

• Insert Size: The Ki67 CDS is exceptionally large (approx. 9-10 kb for human), resulting in a total plasmid size often exceeding 15 kb. Use BioVector? high-efficiency transfection reagents optimized for large constructs.

• Structural Integrity: Large DNA sequences are prone to recombination during cloning. It is essential to perform restriction enzyme digestion mapping after plasmid prep to confirm the integrity of the BioVector? Ki67 construct.

Note: For optimal expression levels, ensure the host cells are in an active growth phase before transfection. Consistently use BioVector? LB Medium and BioVector? transfection grade plasmid prep kits to maintain high DNA purity.

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

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