■ 基本信息
啟動(dòng)子: | CMV |
復(fù)制子: | pUC |
終止子: | SV40 poly(A) signal |
質(zhì)粒分類: | 哺乳系列質(zhì)粒；哺乳熒光質(zhì)粒；哺乳藍(lán)色質(zhì)粒 |
質(zhì)粒大小: | 4731bp |
質(zhì)粒標(biāo)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(dá)宿主: | 293T等哺乳細(xì)胞 |
誘導(dǎo)方式: | 無須誘導(dǎo)，瞬時(shí)表達(dá) |
5'測序引物: | CMV-F（CGCAAATGGGCGGTAGGCGTG） |
3'測序引物: | Sv40-polyA-R（GAAATTTGTGATGCTATTGC） |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 哺乳細(xì)胞 |
質(zhì)粒用途: | 蛋白表達(dá) |
片段類型: | ORF
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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(biāo)記: |
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■ 質(zhì)粒簡介
pEBFP-C1是一個(gè)哺乳細(xì)胞藍(lán)色熒光表達(dá)質(zhì)粒，CMV啟動(dòng)目的基因和藍(lán)色熒光基因EBFP融合表達(dá)，在熒光顯微鏡下，可以觀察到藍(lán)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(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請(qǐng)下載：
ZK455pEBFP-C1哺乳熒光質(zhì)粒.txt
