■ 基本信息
啟動子: | CMV |
復制子: | pUC |
終止子: | SV40 poly(A) signal |
質粒分類: | 哺乳系列質粒；哺乳熒光質粒；哺乳藍色質粒 |
質粒大小: | 4731bp |
質粒標簽: | N-EBFP |
原核抗性: | Kan |
真核抗性: | G418 |
克隆菌株: | DH5a |
培養(yǎng)條件: | 37℃，5%CO2 |
表達宿主: | 293T等哺乳細胞 |
誘導方式: | 無須誘導，瞬時表達 |
5'測序引物: | CMV-F（CGCAAATGGGCGGTAGGCGTG） |
3'測序引物: | Sv40-polyA-R（GAAATTTGTGATGCTATTGC） |
■ 質粒屬性
質粒宿主: | 哺乳細胞 |
質粒用途: | 蛋白表達 |
片段類型: | ORF
|
片段物種: |
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原核抗性: | Kan |
真核抗性: | G418
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熒光標記: |
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■ 質粒簡介
pEBFP-C1是一個哺乳細胞藍色熒光表達質粒，CMV啟動目的基因和藍色熒光基因EBFP融合表達，在熒光顯微鏡下，可以觀察到藍色熒光。
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 .
■ 質粒圖譜
■ 質粒序列
質粒序列請下載：
ZK455pEBFP-C1哺乳熒光質粒.txt
