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- XYbscience
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- XY1142
- 2025年07月10日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
常温
- 保质期：
三年
- 英文名：
pCMS-EGFP
- 库存：
60
- 供应商：
信裕生物
- 规格：
5ug质粒
基本信息
| 质粒类型: | 荧光蛋白报告载体 |
|---|---|
| 启动子: | CMV |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 5541 bp (查看载体序列) |
| 载体标签: | EGFP |
| 载体抗性: | Ampicillin (氨苄青霉素) |
订购信息
| 产品编号 | 产品名称 | 规格 | 价格 |
|---|---|---|---|
| XY1142 | pCMS-EGFP | 5ug质粒 |
¥1000.00 |
质粒图谱
载体描述
pCMS-EGFP is a mammalian expression vector that allows you to express your gene of interest at high levels by cloning it into a multiple cloning site (MCS) downstream of the immediate early promoter of cytomegalovirus (PCMV IE). The intervening sequence (IVS) between PCMV IE and the MCS is an intron that is efficiently spliced out following transcription. SV40 polyadenylation signals downstream of the MCS direct proper processing of the 3' end of the mRNA from your gene of interest. Bacteriophage T7 and T3 promoters are located upstream and downstream of the MCS, respectively.
pCMS-EGFP uses the enhanced green flourescent protein (EGFP) as a transformation marker. EGFP is a red-shifted variant of wild-type GFP from Aquorea victoria (1–3). EGFP has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm; emission maximum = 507 nm.) EGFP encodes the GFPmut1 variant (4) which contains the double-amino-acid substitution of Phe-64 to Leu and Ser-65 to Thr. The coding sequence of the EGFP gene contains more than 190 silent base changes which correspond to human codon-usage preferences (5). Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site (6) to further increase the translation efficiency in eukaryotic cells. EGFP is expressed from the SV40 enhancer/promoter, and a polyadenylation signal from the bovine growth hormone (BGH) gene directs proper processing of the 3' end of the EGFP mRNA. The SV40 origin also allows for replication in mammalian cells expressing the SV40 T antigen.The vector backbone also contains the β-lactamase gene for ampicillin resistance and a ColE1 origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
The name of the vector comes from the order of four critical elements: the CMV IE promoter; the MCS; the SV40 promoter; and EGFP.
载体应用
Genes cloned into the MCS must contain an ATG start codon. pCMS-EGFP and derivatives can be introduced into mammalian cells by any standard method. Transfected cells can be monitored and/or selected by flow cytometry based on the fluorescence of EGFP. Sense or antisense RNA can be transcribed from the T7 and T3 promoters, respectively.
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文献和实验-MEM、DMEM培养基和M-MLV逆转录酶；小牛血清；G418；DIG High PrimeDNA Labeling and Detection Starter Kit I1。 质粒 pGL3-promoter；含EBVR的质粒p220.2。 重组质粒载体pcDNA3-EGFP-EBVR的构建 采用StuI和XbaI消化p220.2，获得长4.7 kb的EBVR，包括EBV核抗原1(EBNA1)和OriP，克隆至pGL3-promotor的SV40启动子下游Hind Ill
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
sixiuwen123 请问我享用流式测凋亡,可现在我的载体本身有绿色荧光,EGFP的表达,本来想用annexin的,现在我该用什么方法上流式 测凋亡呢 freecell 测定凋亡的方法太多了.不一定要用流式. http://gene.bjmu.edu.cn/news/apoptosis.htm 4trggg tunel可以 imhkq
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