■ 基本信息
启动子: | CMV |
复制子: | pUC |
终止子: | SV40 poly(A) signal |
质粒分类: | 哺乳系列质粒；哺乳荧光质粒；哺乳蓝色质粒 |
质粒大小: | 4731bp |
质粒标签: | N-EBFP |
原核抗性: | Kan |
真核抗性: | G418 |
克隆菌株: | DH5a |
培养条件: | 37℃，5%CO2 |
表达宿主: | 293T等哺乳细胞 |
诱导方式: | 无须诱导，瞬时表达 |
5'测序引物: | CMV-F（CGCAAATGGGCGGTAGGCGTG） |
3'测序引物: | Sv40-polyA-R（GAAATTTGTGATGCTATTGC） |
■ 质粒属性
质粒宿主: | 哺乳细胞 |
质粒用途: | 蛋白表达 |
片段类型: | ORF
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片段物种: |
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原核抗性: | Kan |
真核抗性: | G418
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荧光标记: |
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■ 质粒简介
pEBFP-C1是一个哺乳细胞蓝色荧光表达质粒，CMV启动目的基因和蓝色荧光基因EBFP融合表达，在荧光显微镜下，可以观察到蓝色荧光。
pEBFP-C1 carries a blue fluorescent variant of the Aequorea victoria green fluorescent protein gene (GFP). The EBFP gene contains four amino acid substitutions. The Tyr-66 to His substitution gives EBFP fluorescence excitation and emission maxima (380 and 440 nm, respectively) similar to other blue emission variants. The other three substitutions (Phe-64 to Leu; Ser-65 to Thr; and Tyr- 145 to Phe) enhance the brightness and solubility of the protein, primarily due to improved proteinfolding properties and efficiency of chromophore formation. The Em of EBFP is 31,000 cm–1M–1 for 380-nm excitation, leading to a fluorescent signal that is 2–3-fold brighter than other blue variants of GFP and roughly equivalent to wt GFP. In addition, the rate of photobleaching of EBFP is one-half to one-third that of P4-3, a popular predecessor to EBFP. EBFP contains >190 silent mutations that create an open reading frame comprised almost entirely of preferred human codons .
■ 质粒图谱
■ 质粒序列
质粒序列请下载：
ZK455pEBFP-C1哺乳荧光质粒.txt
