蛋白質(zhì)及其標(biāo)記ppt課件(編輯修改稿)
2025-02-15 01:39 本頁(yè)面
【文章內(nèi)容簡(jiǎn)介】 Labeling with Green Fluorescent Protein (GFP) GFP (or the newer EGFP) is a 27 kD (236 residue) protein isolated from the jellyfish Aequorea victoria. The structure consists of an 11stranded bbarrel surrounding a single central strand. After the protein folds, three residues spontaneously cyclize in an unprecedented fashion. A subsequent oxidation (nonenzymatic) results in the formation of a strong fluorescent chromophore. Although the wt protein exhibits green fluorescence, blue, cyan, and red mutants have now been identified. The chromophore does not fluoresce when removed from the protein core. 1. Protein folding 2. Cyclization 3. Oxidation Roger Tsien UCSD 雙砷染料 四半胱氨酸體系 與連載重組蛋白質(zhì)末端的六肽標(biāo)簽 CCXXCC序列（ TCtag）特異性標(biāo)記 Roger Tsien UCSD ? 雙砷染料 四半胱氨酸體系的熒光強(qiáng)度是雙砷染料的 50000 倍。 ? 跟熒光蛋白相比，四半胱氨酸標(biāo)簽序列體積小，不會(huì)影響被標(biāo)記蛋白質(zhì) 或細(xì)胞的正常生理功能，另外染料量子產(chǎn)率高、穩(wěn)定性強(qiáng)、標(biāo)記速度快 OO O HC O 2 HA s A sS S S SNOO O HA s A sS S S SOH O O HA s A sS S S SOC lC lReAsH (a phenoxaine derivative) CHOxAsH (a dihydroxyxanthone derivative) Tsien et al. Science 1998, 281, 269271. and JACS 2022, 124, 60636076. 雙砷染料 四半胱氨酸體系 雙砷染料 四半胱氨酸重組噬菌體的構(gòu)建及 應(yīng)用 Angew. Chem. Int. Ed., 2022, 50, 58735877 其他特殊標(biāo)記方法： NTA熒光團(tuán)綴合物探針 ?由兩部分組成，一部分為熒光基團(tuán)，另一部分為螯合物， 由金屬離子與含有三醋酸根的叔胺類(lèi)化合物組成（ NTA） ?六組氨酸 (His6 ) 識(shí)別結(jié)合 Ni2+：次氮基三乙酸（ Ni2+:NTA) ? 蛋白質(zhì) C端、 N端、或肽鏈中間 ?比雙砷染料 四半胱氨酸體系通用 The term ‘‘click chemistry’’, originally brought up by K. Barry Sharpless at The Scripps Research Institute in 2022. K. Barry Sharpless and his coworkers have discovered and developed many widely used catalytic oxidation processes, the Sharpless reactions for asymmetric epoxidation, dihydroxylation, and aminohydroxylation of olefins （烯烴的不對(duì)稱(chēng)環(huán)氧化、雙羥化和羥氨化反應(yīng)） . The Nobel Prize, 2022 點(diǎn)擊化學(xué) Azide/Alkyne Cycloaddition L. Pauling. Proc. Natl. Acad. Sci. USA 1933, 19, 860867。 Huisgen, R. Angew. Chem. Int. Ed. 1963, 2, 633696 Sharpless, . et al. Angew. Chem. Int. Ed 2022, 41, 25962599。 Meldal,. et al. J. Org. Chem. 2022, 67, 30573064 R 39。39。 R 39。N 3 N NN R 39。R 39。39。15+8 0 o C N N
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