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引用和文献 (2)
Invitrogen™
pLenti6/V5 Directional TOPO™ Cloning Kit
The pLenti6⁄V5 Directional TOPO™ Cloning Kit contains the TOPO™-adapted ViraPower™ lentiviral expression vector, pLenti6⁄V5-D-TOPO™ for quick PCR-based cloning and high-level了解更多信息
| 货号 | 数量 |
|---|---|
| K495510 | 20 Reactions |
货号 K495510
价格（CNY)
36,120.00
Each
数量:
20 Reactions
价格（CNY)
36,120.00
Each
The pLenti6⁄V5 Directional TOPO™ Cloning Kit contains the TOPO™-adapted ViraPower™ lentiviral expression vector, pLenti6⁄V5-D-TOPO™ for quick PCR-based cloning and high-level expression of a target gene in dividing and non-dividing mammalian cells. The vector has the CMV promoter for driving high-level, constitutive expression of the target gene and the blasticidin selection marker for stable selection in mammalian cells.
Advantages
• High efficiency and rapid cloning
• Constitutive gene expression in dividing and non-dividing mammalian cells in vitro or in vivo
• Produces replication-incompetent virus for enhanced biosafety of the system
Key Features
• Directional TOPO™ Cloning site for rapid and efficient directional cloning of blunt-end PCR products
• Rouse Sarcoma Virus (RSV) enhancer⁄promoter for Tat-independent production of viral mRNA in the producer cell line
• Modified HIV-1 5’ and 3’ Long Terminal Repeats (LTR) for viral packaging and reverse transcription
• HIV-1 psi (ψ) packaging sequence for viral packaging
• HIV Rev response element (RRE) for Rev-dependent nuclear export of unspliced viral mRNA
• (CMV) immediate early promoter for high-level constitutive expression of the gene of interest in mammalian cells
• C-terminal V5 epitope for detection of the recombinant protein
• Blasticidin (bsd) resistance gene for selection in E. coli and mammalian cells
• Ampicillin resistance gene for selection in E. coli
• pUC origin for high-copy replication and maintenance of the plasmid in E. coli
Kit includes
pLenti6⁄V5-D- TOPO™ Reagents
One Shot™ Stbl3™ Chemically Competent E. coli
For research use only. Not intended for any therapeutic or diagnostic use.
Advantages
• High efficiency and rapid cloning
• Constitutive gene expression in dividing and non-dividing mammalian cells in vitro or in vivo
• Produces replication-incompetent virus for enhanced biosafety of the system
Key Features
• Directional TOPO™ Cloning site for rapid and efficient directional cloning of blunt-end PCR products
• Rouse Sarcoma Virus (RSV) enhancer⁄promoter for Tat-independent production of viral mRNA in the producer cell line
• Modified HIV-1 5’ and 3’ Long Terminal Repeats (LTR) for viral packaging and reverse transcription
• HIV-1 psi (ψ) packaging sequence for viral packaging
• HIV Rev response element (RRE) for Rev-dependent nuclear export of unspliced viral mRNA
• (CMV) immediate early promoter for high-level constitutive expression of the gene of interest in mammalian cells
• C-terminal V5 epitope for detection of the recombinant protein
• Blasticidin (bsd) resistance gene for selection in E. coli and mammalian cells
• Ampicillin resistance gene for selection in E. coli
• pUC origin for high-copy replication and maintenance of the plasmid in E. coli
Kit includes
pLenti6⁄V5-D- TOPO™ Reagents
One Shot™ Stbl3™ Chemically Competent E. coli
For research use only. Not intended for any therapeutic or diagnostic use.
仅供科研使用。不可用于诊断程序。
规格
克隆方法定向 TOPO™
构成或诱导系统Constitutive
输送类型Lentiviral
适用于（应用）Viral Expression
反应次数20 reactions
产品线TOPO, ViraPower
产品类型TOPO 克隆试剂盒
促进剂CMV
蛋白标记V5 抗原决定簇标签
数量20 Reactions
选择试剂（真核生物）Blasticidin
载体pLenti, Directional TOPO Vectors
Unit SizeEach
内容与储存
pLenti⁄V5-D-TOPO™ reagents (Box 1) Store at -20°C
• pLenti6⁄V5-D-TOPO™ Reagents (15-20 ng⁄μl linearized plasmid DNA in: 50% glycerol, 50 mM Tris-HCl, pH 7.4 (at 25°C),1 mM EDTA, 2 mM DTT, 0.1% Triton X-100, 100 μg⁄ml BSA, 30 μM bromophenol blue)
• dNTP Mix (12.5 mM each dATP, dCTP, dGTP, dTTP in water, pH8 at a total volume of 10 μl)
• Salt Solution (1.2 M NaCl and 0.06 M MgCl2, volume of 50 μl)
• Water (1 ml)
• CMV Forward Sequencing Primer (0.1 μg⁄μl in TE Buffer, pH 8, pH8 at a total volume of 20 μl)
• Control PCR Primers (0.1 μg⁄μl each in TE Buffer, pH 8, pH8 at a total volume of 10 μl)
• Control PCR Template (0.1 μg⁄μl in TE Buffer, pH 8, pH8 at a total volume of 10 μl)
• pLenti6⁄V5-GW⁄lacZ (lyophilized in TE Buffer, pH 8, 10 μg)
One Shot™ Stbl3™ Chemically Competent E. coli (Box 2) Store at -80°C
• S.O.C. Medium (2% Trypton, 0.5% Yeast Extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl2, 10 mM MgSO4, 20 mM glucose. 6ml) May be stored at +4°C or room temperature
• Stbl3™ Cells (21 x 50 μl)
• pUC19 Control DNA (10 pg⁄μl in 5 mM Tris-HCl, 0.5 mM EDTA, pH 8. 50 μl)
• pLenti6⁄V5-D-TOPO™ Reagents (15-20 ng⁄μl linearized plasmid DNA in: 50% glycerol, 50 mM Tris-HCl, pH 7.4 (at 25°C),1 mM EDTA, 2 mM DTT, 0.1% Triton X-100, 100 μg⁄ml BSA, 30 μM bromophenol blue)
• dNTP Mix (12.5 mM each dATP, dCTP, dGTP, dTTP in water, pH8 at a total volume of 10 μl)
• Salt Solution (1.2 M NaCl and 0.06 M MgCl2, volume of 50 μl)
• Water (1 ml)
• CMV Forward Sequencing Primer (0.1 μg⁄μl in TE Buffer, pH 8, pH8 at a total volume of 20 μl)
• Control PCR Primers (0.1 μg⁄μl each in TE Buffer, pH 8, pH8 at a total volume of 10 μl)
• Control PCR Template (0.1 μg⁄μl in TE Buffer, pH 8, pH8 at a total volume of 10 μl)
• pLenti6⁄V5-GW⁄lacZ (lyophilized in TE Buffer, pH 8, 10 μg)
One Shot™ Stbl3™ Chemically Competent E. coli (Box 2) Store at -80°C
• S.O.C. Medium (2% Trypton, 0.5% Yeast Extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl2, 10 mM MgSO4, 20 mM glucose. 6ml) May be stored at +4°C or room temperature
• Stbl3™ Cells (21 x 50 μl)
• pUC19 Control DNA (10 pg⁄μl in 5 mM Tris-HCl, 0.5 mM EDTA, pH 8. 50 μl)
常见问题解答 (FAQ)
我进行了稳转筛选，但我的抗生素耐受克隆中未表达我的目的基因。发生了什么问题？
我正在使用一款哺乳动物表达载体，但未成功表达我的蛋白。你们能帮我解决这一难题么？
我正在使用一个包含新霉素抗性基因的哺乳动物表达载体。我能否在哺乳动物细胞中使用新霉素进行稳转筛选？
我构建的载体中，目的基因的ATG前方还有另一个ATG，这样可以么？它会干扰我基因的翻译么？
你们是否提供表达GFP的哺乳动物载体，这样我就可将其作为参照来监测我的转染和表达情况？
引用和文献 (2)
引用和文献
Abstract
RIP140-targeted repression of gene expression in adipocytes.
Journal:Mol Cell Biol
PubMed ID:16227589
'Ligand-dependent repression of nuclear receptor activity forms a novel mechanism for regulating gene expression. To investigate the intrinsic role of the corepressor RIP140, we have monitored gene expression profiles in cells that express or lack the RIP140 gene and that can be induced to undergo adipogenesis in vitro. In contrast
Coregulation of vascular tube stabilization by endothelial cell TIMP-2 and pericyte TIMP-3.
Journal:J Cell Biol
PubMed ID:17030988
The endothelial cell (EC)-derived tissue inhibitor of metalloproteinase-2 (TIMP-2) and pericyte-derived TIMP-3 are shown to coregulate human capillary tube stabilization following EC-pericyte interactions through a combined ability to block EC tube morphogenesis and regression in three-dimensional collagen matrices. EC-pericyte interactions strongly induce TIMP-3 expression by pericytes, whereas ECs produce TIMP-2