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- 详细信息
- 文献和实验
- 技术资料
- 库存：
60
- 英文名：
pLVX-EF1α-IRES-Puro
- 保质期：
三年
- 供应商：
再康生物
- 保存条件：
常温
- 规格：
5ug质粒
基本信息
| 质粒类型: | 慢病毒载体 |
|---|---|
| 高拷贝/低拷贝: | 高拷贝 |
| 启动子: | EF1α/ EF1a |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 8825 bp (查看载体序列) |
| 5' 测序引物及序列: | EF-1a Forward: TCAAGCCTCAGACAGTGGTTC |
| 3' 测序引物及序列: | IRES-R: CCTCACATTGCCAAAAGACG |
| 载体抗性: | Ampicillin (氨苄 青 霉 素) |
| 筛选标记: | 嘌呤霉素(Puromycin) |
| 备注: | 含有IRES序列和EF1a启动子，慢病毒表达载体 |
订购信息
| 产品编号 | 产品名称 | 规格 | 价格 |
|---|---|---|---|
| ZK2015 | pLVX-EF1α-IRES-Puro | 5ug质粒 |
¥1500.00 |
质粒图谱
载体描述
pLVX-EF1α-IRES-Puro is an HIV-1-based, lentiviral expression vector designed to simultaneously and constitutively express a protein of interest and puromycin resistance (Puror) from a bicistronic transcript in mammalian cells. Expression of your protein of interest and Puror from a bicistronic transcript allows puromycin resistance to be used as an indicator of transduction efficiency and a marker for selection.
Simultaneous expression of a protein of interest and puromycin resistance is made possible by the presence of an encephalomyocarditis virus internal ribosome entry site (IRES; 1) positioned between the multiple cloning site (MCS) and Puror. The IRES allows a protein of interest and puromycin resistance to be translated from a single bicistronic mRNA. Stable, constitutive expression of the bicistronic transcript is driven by the EF1α promoter (PEF1α), which continues to be constitutively active even after vector integration into the host cell genome (2).
pLVX-EF1α-IRES-Puro contains all of the viral processing elements necessary for the production of replicationincompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral RNA (3), leading to increased viral titers from packaging cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (4). Finally, pLVX-EF1α-IRES-Puro also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (5). The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.
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naj2007 请教一下各位老师，在构建过表达载体的时候，VEGF和GFP是不是不可以做成融合蛋白，但是如果用不同的启动子的话，很容易丢失荧光，有没有什么解决方法。 zhujoker 可以做成融合蛋白的，即使是使用不同的启动子的话荧光也不那么容易丢失的，大不了你将荧光的启动子换成强的如CMV，EF1a之类。 kinguangjun 或者你可以用CMV-VEGF-IRES-GFP来做
但感觉自己包装效率还是不太高，做起来不好弄 kinguangjun 你可以考虑clontech的pLVX-ShRNA2和pLVX-IRES-ZsGreen1慢病毒系统，比invitrogen的慢病毒系统要高两个数量级以上，我们实验室一直都用这个系统。 jinanfendou 看你做什么实验了，慢病毒载体系统很多公司有卖的，但是你自己包装的话，滴度可能达不到这么高，好转染的细胞还是可以的，但是要是难转染的细胞和动物
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