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网站首页 >> pEBFP-N1哺乳荧光质粒 货号：P2115
pEBFP-N1哺乳荧光质粒 货号：P2115

信息

启动子 CMV
复制子 pUC
终止子 SV40 poly(A) signal
质粒分类 哺乳细胞，荧光蛋白报告载体
原核抗性 Kan
真核抗性 G418
克隆菌株 DH5α
培养条件 37℃
表达宿主 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 根据序列设计引物
质粒宿主 哺乳细胞
质粒用途 蛋白表达
片段类型 基因CDS
原核抗性 卡那霉素

简介
     pEBFP-N1 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 (1–3). 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 (1, 4, 5). 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 (1). EBFP contains >190 silent mutations that create an open reading frame comprised almost entirely of preferred human codons (6). Furthermore, upstream sequences flanking EBFP have been converted to a Kozak consensus translation initiation site (7). These changes increase the translational efficiency of the EBFP mRNA and consequently the expression of EBFP in mammalian and plant cells.


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