BioVector® Ki67 过表达质粒 / BioVector® Ki67 Overexpression Plasmid

通用定义 / General Definition:

BioVector® Ki67 过表达质粒是一种专门设计的真核表达载体，用于在哺乳动物细胞中高效表达 Ki67（由 MKI67 基因编码） 蛋白。Ki67 是一种关键的增殖标记蛋白，仅在细胞周期的活跃阶段（G1、S、G2 和 M 期）表达，而在静止期（G0 期）缺失。该过表达模型广泛用于研究细胞增殖调控、肿瘤进展机制、细胞周期动力学以及作为癌症预后研究的阳性对照。

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 过表达质粒技术说明书 (Plasmid Datasheet)

中文版说明书

1. 产品基本信息

• 产品名称： BioVector® Ki67 过表达质粒

• 目的基因： 人/小鼠 MKI67 (Ki67)

• 启动子： BioVector® CMV 或 EF1$\alpha$ 强启动子

• 载体骨架： BioVector® pCDNA3.1(+) 或类似哺乳动物表达骨架

• 抗性基因： * 大肠杆菌： 氨苄青霉素 (Ampicillin) 或 卡那霉素 (Kanamycin)

  • 真核筛选： 新霉素 (G418) 或 嘌呤霉素 (Puromycin)

• 标签： 可选 Flag, Myc, 或 GFP 标签（用于蛋白定位与检测）

2. 培养与扩增条件

• 宿主菌： BioVector® DH5$\alpha$ 或 Stbl3（针对 Ki67 这种大片段基因建议使用高稳定性菌株）。

• 培养基： BioVector® LB 培养基

• 抗生素添加： 按载体要求添加 BioVector® 氨苄青霉素 (100 $\mu$g/mL)。

• 培养环境： 37°C 恒温振荡培养。

3. 载体应用

• 细胞增殖研究： 诱导静止期细胞进入细胞周期。

• 肿瘤模型构建： 研究 Ki67 高表达对肿瘤生长和侵袭的影响。

• 抗体检测验证： 作为 Ki67 免疫组化 (IHC) 或流式细胞术 (FACS) 检测的阳性对照。

4. 注意事项

• 片段大小： Ki67 编码区序列非常大（人源约 9-10 kb），质粒总长可能超过 15 kb，转染时建议使用 BioVector® 高效大片段转染试剂。

• 序列稳定性： 大片段 DNA 在克隆过程中易发生重组，建议扩增后进行双酶切鉴定。

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生产厂家: BioVector NTCC典型培养物保藏中心

E-mail: BioVector@163.com

http://www.biovector.net