| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pISRE-TA-Luc |
| 质粒类型: | 信号通路报告系统载体 |
| 高拷贝/低拷贝: | -- |
| 启动子: | -- |
| 克隆方法: | 多克隆位点,限制性内切酶 |
| 载体大小: | 4.9kb |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | -- |
| 载体抗性: | Ampicillin |
| 筛选标记: | -- |
| 备注: | -- |
| 产品目录号: | ZY631915 |
| 稳定性: | -- |
| 组成型: | -- |
| 病毒/非病毒: | -- |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃低温保存
- 保质期:
三年
- 英文名:
pISRE-TA-Luc
- 库存:
20
- 供应商:
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息


载体简介
pISRE-TA-Luc is designed for monitoring the induction of the STAT1 and STAT2 components of Jak/STAT-mediated signal
transduction pathways. Signaling molecules, including type I(IFN-α and -β) and type II (IFN-γ) interferons (1), induce
signalling by binding receptors and causing receptor dimerization at the cell surface. This dimerization causes the receptor
itself to be phosphorylated and act as a docking site for transcription factors, including STAT1 and STAT2. The STAT proteins
are then phosphorylated, dimerize and translocate to the nucleus, where the STAT1 and STAT2 heterodimer regulates transcription by binding to the IFN-stimulated response element (ISRE; 2). pISRE-TA-Luc contains five copies of the ISRE enhancer element,
located upstream of the minimal TA promoter, the TATA box from the herpes simplex virus thymidine kinase promoter (PTA).
Located downstream of the PTA is the firefly luciferase gene (luc). Upon binding of the STAT1 and STAT2 heterodimer to the
cis-acting ISRE enhancer element, transcription is induced and the reporter gene is activated.
The luciferase coding sequence is followed by the SV40 late polyadenylation signal to ensure proper, efficient processing of
the luciferase transcript in eukaryotic cells. A synthetic transcription blocker (TB) is located upstream of the cis-acting
enhancer element. It is composed of adjacent polyadenylation and transcription pause sites for blocking nonspecific
transcription (3). The vector backbone also contains an f1 origin for single-stranded DNA production, a pUC origin of
replication, and an ampicillin resistance gene for propagation and selection in E. coli.
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文献和实验静脉注射,病毒注射总量为 1×10^11 vg/小鼠。Luc 的表达由 cTnT 启动子控制。(DOI: 10.1038/gt.2010.105) 图 4. 五种 AAV 血清型在小鼠体内的心脏基因递送效率比较。注射方式为左心室前壁注射,病毒注射总量为 5×10^10 vg/小鼠,体积为 10μL。Luc 的表达由 cTnT 启 动子控制。(DOI:10.1002/jgm.1576) 图 5. 五种 AAV 血清型以不同剂量注射至小鼠体内的心脏基因递送效率比较。注射方式为颈静脉注射。EGFP
3`-T突出端的载体用于直接高效地连接PCR产物。系统中也包含感受态细胞和S.O.C培养基, 使得实验操作更为简单方便。所以TA克隆不需使用含限制酶序列的引物,不需将PCR产物进行优化,不需把PCR产物做平端处理(Invitrogen另有平端载体供高确限度酶克隆)。不需在PCR扩增产物上加接头,即可直接进行克隆。图32 The TA Cloning® ConceptTopo TA克隆方法(Topo TA Cloning Kit)Topo TA克隆原理与TA克隆一样,唯一不同的是TA克隆用的是T
) 利用Taq聚合酶同时具有的末端连接酶的功能,在每条PCR 扩增产物的3`端自动添加一个3`-A突出端。Invitrogen公司TA克隆 系统和Topo TA克隆 系统都提供一个线性含3`-T突出端的载体用于直接高效地连接PCR 产物。系统中也包含感受态细胞和S.O.C培养基, 使得实验操作更为简单方便。 所以TA克隆 不需使用含限制酶序列的引物,不需将PCR 产物进行优化,不需把PCR 产物做平端处理(Invitrogen另有平端载体供高确
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