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- LSM Bio
- 进口
- PVT10821
- 2025年08月05日
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- 技术资料
- 保存条件：
负20摄氏度
- 保质期：
3个月
- 英文名：
pGRE-luc
- 库存：
50
- 供应商：
LSMBIO
- 规格：
2ug
pGRE-luc
PVT10821 2ug
pGRE-luc Description
PGRE-Luc can be used to detect the activity of GRE signaling pathway. Carrier pGRE-Luc contains luciferase gene from firefly. The vector also contains multiple copies of the GRE binding site sequence, which is fused with a TATA-like promoter (PTAL) derived from the herpes simplex virus thymidine kinase (HSV-TK) promoter. When endogenous GRE proteins bind to GRE enhancers, transcription begins and the reporter gene begins to express. The luciferase encoded sequence was followed by a SV40 polyadenylate signal to ensure correct and efficient transcription in eukaryotic cells. There is a synthetic transcription inhibitory region (TB) upstream of the GRE binding site, which is composed of polyadenylate signaling and transcription termination sites to reduce the transcriptional background. The vector skeleton also contains an F1 replication initiation region for single strand DNA replication, a pUC replication initiation site and an ampicillin resistance gene for culture and screening.
The vector pGRE-Luc is used to detect the interaction between transcription factor GRE and GRE enhancer, providing a direct way to detect the activity of this pathway. The vector can be co-transfected with a target gene expression vector to observe whether it affects the activity of this pathway. Luciferase is a highly sensitive proenzyme reporter protein, suitable for any standard luciferase assay, can be used to quantify the level of expression. The pGRE-Luc vector can be transfected into mammalian cells by any standard method. In order to screen stable clones, transfected vectors usually contain resistant screening genes such as neomycin, hygromycin or puromycin.
The suitable host system is DH5 alpha and some other common lines. The single strand DNA synthesis of host bacteria requires a F 'episome, such as JM109. Screening marker: The plasmid provides the starting site of E. coli replication with ampicillin resistance (100ug/ml): pUC copy number: ~500 incompatible plasmid: pMB1/ColE1.
pGRE-luc Informaiton
Replicon: pUC
Prokaryotic resistance: ampicillin Amp
Screening marker: luciferase Luc
Clone strain: Escherichia coli DH5 alpha
Culture conditions: 37 鈩?/p>
pGRE-luc Multiple cloning site
pGRE-luc Sequence
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文献和实验superskyfly 我自己都觉得我自己很牛，千百人都做出来的试验，我就是做不出来。质粒用的是Promega p4.32-NF-kB-Luc, 转染用的是Invitrogen的Lipofactamin, 激活用的是Sigma生产的PMA和PHA-P,Assay用的是Promega dual,结果加了PMA和PHA的和不加的读数几乎没有区别。加了compound的和不加的也没有区别。 用过PC3, Jurkat, A549，HCT116都是一样。
tianjiaoya 我想构建一个真核表达质粒，用这个真核表达质粒来表达绿色荧光蛋白，但是我想让绿色荧光蛋白的表达受酵母菌GLA4转录因子的调控，也就是说有GLA4存在，这个质粒就表达绿色荧光蛋白，没有GLA4存在，就不表达绿色荧光蛋白。当然需要对真核表达质粒上的启动子做相应的调整，但是我现在的问题是，转录因子GLA4能不能对质粒上有相应调控元件的表达盒，进行调控？据说GLA4上有核定位元件，要是将这个核定位序列删除，可行吗？据说核内有很多的辅助因子，参与了蛋白
问：我要做LUCIFERASE 了，可是我一点概念都没有。有哪位有具体的操作PROTOCOL？还有一些白痴问题luciferase 要检测什么呢？是把我的带有LUC的质粒转染到细胞里，然后检测它的活性？什么意思啊？我检测它的目的是什么呢？我只知道我现在有一个LUC的质粒，老板让我准备做，我都不知道是什么目的和思路......谢谢啦答：luciferase检测的是虫荧光素酶基因的表达情况，一般我们构建luciferase载体的时候是把待研究的基因装在luciferase基因的前面，从而以
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