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- 2025年12月29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：
pECFP-C1
- 库存：
现货
- 供应商：
上海盖宁生物
| 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: | 4731 bp |
| 5' 测序引物及序列: | EGFP-C: 5'd[CATGGTCCTGCTGGAGTTCGTG] |
| 载体标签: | ECFP (Nterm) |
| 载体抗性: | Kanamycin (卡那霉素) |
| 筛选标记: | Neomycin (新霉素) |
订购信息
| 产品编号 | 产品名称 | 规格 | 价格 |
|---|---|---|---|
| 1114 | pECFP-C1 | 5ug质粒 |
¥1000.00 |
质粒图谱
载体描述
pECFP-C1 encodes an enhanced cyan fluorescent variant of the Aequorea victoria green fluorescent protein gene (GFP). The ECFP gene contains six amino acid substitutions. The Tyr-66 to Trp substitution gives ECFP fluorescence excitation (major peak at 433 nm and a minor peak at 453 nm) and emission (major peak at 475 nm and a minor peak at 501 nm) similar to other cyan emission variants (1–3). The other five substitutions (Phe-64 to Leu; Ser-65 to Thr; Asn-146 to Ile; Met-153 to Thr; and Val-163 to Ala) enhance the brightness and solubility of the protein, primarily due to improved protein-folding properties and efficiency of chromophore formation (2, 4, 5).
In addition to the chromophore mutations, ECFP contains >190 silent mutations that create an open reading frame comprised almost entirely of preferred human codons (6). Furthermore, upstream sequences flanking ECFP have been converted to a Kozak consensus translation initiation site (7). These changes increase the translational efficiency of the ECFP mRNA and consequently the expression of ECFP in mammalian and plant cells.
The MCS in pECFP-C1 is between the ECFP coding sequence and the stop codon. Genes cloned into the MCS will be expressed as fusions to the C-terminus of ECFP if they are in the same reading frame as ECFP and there are no intervening in-frame stop codons. ECFP with a C-terminal fusion moiety retains the fluorescent properties of the native protein and thus can be used to localize fusion proteins in vivo.
The vector contains an SV40 origin for replication and a neomycin resistance (Neor) gene for selection (using G418) in eukaryotic cells. A bacterial promoter (P) upstream of Neor expresses kanamycin resistance in E. coli. The vector backbone also provides a pUC19 origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
The recombinant ECFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transfectants can be selected using G418 (8). pECFP-C1 can also be used simply to express ECFP in a cell line of interest (e.g., as a transfection marker).
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文献和实验*发表【中文论文】请标注：由上海盖宁生物科技有限公司提供；
*发表【英文论文】请标注：From Shanghai Gaining Biotechnology Co., Ltd.
pECFP-C pEGFP-C-5’ pECFP-C-3' pECFP-N1 pEGFP-N-5’ pEGFP-N-3’ pEF/myc/ER pEF-F pCDNA3.1R pEGFP-C(KAN+) pEGFP-C-5’ pEGFP-C-3' pEGFP-N(KAN+) pEGFP-N-5’ pEGFP-N-3’ pENTR/D-TOPO(KAN+) M13F M13R pET-*(His)(KAN+) T7 T7 Ter pET22(a,b,c)(AMP+) T7 T7 Ter
RpEASY-BluntSimpleM13FT7/ M13R（距离插入位点合适，优先安排）pEASY-T1 SimpleM13FM13R（距离插入位点合适，优先安排）pEASY-T1M13F/T7M13RpEASY-T3M13F/T7M13R/SP6pECFP-N(C)PECFP-N-5.PECFP-N-3,pEF/myc/cyto/ER/mito/nucpEF-5Pcdna3.1R/BGH revpEF1(4,6/myc-His)T7/pEF-5Pcdna3.1R/BGH revpEF1(4,6/V
. coli/mammals pECFP-C1 pECFPN1 表达 青色 荧光蛋白和目的基因的融合蛋白，目的基因位于N端 kan/neo 4.7kb E. coli/mammals pEYFP-C1 pEYFPC1 表达 黄色 荧光蛋白和目的基因的融合蛋白，目的基因位于C端 kan/neo 4.7kb E. coli/mammals 4)、哺乳动物/真核荧光素酶载体（研究启动子专用）来自荧火虫的化学发光剂，加入底物后发光，不适合追踪单个细胞。 PGL3-basic PGL
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