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元件指南
VectorBuilder提供了許多常用的載體元件，可供用物術戶在設計載體時(shí)進(jìn)事下行選擇。下表按類型提供了這(zhè)些常用元件的詳細信息。
表位标簽蛋白
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| FLAG | FLAG epitope tag | Facilitates antibody binding of 道司tagged protein. | J Biochem Biophys Methods. 49:455城答 (2001); Appl Microbiol Biotechno內這l. 60:523 (2003歌遠) | View |
| 3xFLAG | 3 tandem FLAG epitope機湖 tag | Facilitates antibody bi黑讀nding of tagged protein. | Appl Microbiol Biotechnol. 60:523 (2音關003) | View |
| HA | Hemagglutinin epitope tag | Facilitates antibody b國這inding of tagged protein.也技 | Nature. 443:230 做廠(2006) | View |
| Myc | c-Myc epitope tag | Facilitates antibody binding少黃 of tagged protein. | Appl Microbiol Biot化長echnol. 60:523 醫亮(2003) | View |
| V5 | V5 epitope tag | Facilitates antib個不ody binding of t湖舞agged protein. | J Biol Chem. 282:20799 (2007) | View |
| OVA(257-264) | H-2Kb-restricted OVA MHC cla睡鐘ss I epitope | Allows detection of a strong CD8關工+ cytolytic T cell response靜信. | Eur J Immunol. 21:2891 (19請行91); J Immunol. 150:1212了煙 (1993) | View |
| OVA(323-339) | I-Ad-restricted OVA MHC件北 class II epitope | Allows studying the nature of class II 土又MHC-peptide binding and T-cell activ相高ation. | Biochemistry. 38:16663 (1999); J Immunol. 171:1999 (2003) | View |
| 10xGCN4_v4 | The v4 version of the G師工CN4 peptide arra車靜y with 10 copies of the peptide bin靜市ding site, also know as SunTag10x_樹光v4 | Allows tagged proteins to be recogni冷對zed by the GCN4 scFv antibod話愛y. | Cell. 159: 635 (2014你公) | View |
| Rho1D4 | A 9 amino acid 1D4 epitope derived from森又 the C-terminus of bovine rhodops議的in | Allows tagged proteins文歌 to be recognized by the 女但Rho1D4 antibody; typically recom都樹mended for the purification and char相車acterization of membran購睡e proteins. | Methods Mol Biol. 1177:1 (20業術14) | View |
| ALFA-tag | ALFA-tag | Allows tagged proteins to be 土就recognized by the 見去NbALFA nanobody. | Nature Communications. 10:4403匠農 (2019) | View |
純化标簽
| 姓名 | 描述 | Application Note服說s | References | 序列 |
|---|---|---|---|---|
| 6xHis | 6 tandem histidine tag | Facilitates puri子姐fication by nickel 冷科or cobalt matrix and antibody bind子廠ing of tagged protein. | Appl Microbiol Biotechno校一l. 60:523 (2003) | View |
| 10xHis | 10 tandem histidine tag | Facilitates purification by nickel為綠 or cobalt matrix and antib都吧ody binding of tagged protein. | Appl Microbiol Biotechnol又長. 60:523 (2003) | View |
| Strep-tag_II | Strep-tag II | Facilitates purificat子花ion by Strep-Tactin (modifie一子d streptavidin) matr草吧ix and antibody binding of tagged pr頻嗎otein. | Appl Microbiol Biote老通chnol. 60:523 (2003) | View |
| Twin-Strep-tag | 2 tandem Strep-ta雜術g II linked wit在物h 3xGGGS | Facilitates purification by Strep-Tact還呢in (modified streptavidin) m我內atrix and antibody都一 binding of tagged 區微protein. | Protein Expr Purif. 92:54 (2013) | View |
| S-tag | S-tag | Facilitates purification by S-p紙算rotein matrix and antibody binding 內多of tagged protein. | Appl Microbiol Biotechnol. 60:523 (男務2003) | View |
| MBP | Maltose-binding protein | Facilitates purification by am水友ylose matrix and呢呢 antibody binding of tagg看不ed protein; increases th算員e solubility of tagged pr東爸otein in bacteria. | Appl Microbiol Biotech刀舞nol. 60:523 (200事鐘3) | View |
| GST | Glutathione S-transferase | Facilitates purification by glu和國tathione matrix and a朋也ntibody binding of t新個agged protein; helps to stabilize tagg木船ed protein and protect it agains時短t intracellular 月船protease cleavage. | Appl Microbiol Biotechno人不l. 60:523 (2003) | View |
| Avi | Avi tag | Facilitates biotinylation 懂靜of tagged protein by the enzym如大e BirA so the protein can be 輛房isolated by streptavidin. | Protein Sci. 8:921 (1999); Methods Mol Biol. 1266: 171 (2015) | View |
| Halo | HaloTag7 | Increases expres知城sion and solubility of t些低agged protein; enabl她唱es covalent, efficient and specific cap計雨ture of a protein of intere火錯st onto HaloLink™ Resin; high yi司舊eld and purity. | Protein Expr Purif. 68:1嗎行10 (2009) | View |
| Halo-TEV | HaloTag7 linked wi嗎也th TEV site | Increases expression and愛視 solubility of tagged prote學得in; enables covalent, efficient an友路d specific capture of a protei了劇n of interest onto Halo件熱Link™ Resin; high yi男志eld and purity; ca工要n be removed by TEV prote跳腦ase; N-terminal tag. | Protein Expr Purif. 68:110 (2009刀們) | View |
| SUMO | Small ubiquitin-like mo雜影difier tag | Facilitates purification by enhancing s樹老tability and solubility分也 of tagged proteins; can be sp光子ecifically recogn黑相ized by SUMO proteases. | Protein Sci.22(11)身懂:1466-77 (2013) | View |
| hIgG-Fc | The constant re場話gion (CH2 and CH3 domains) of human 暗你immunoglobulin heavy chain | Increases protein yie關們ld; allows rapid and simple d我路etection of protein 雜去expression; enables simple protein G/刀黃A affinity chromatograp就綠hy based purification of Fc-tag土少ged proteins. | EMBO Mol Med. 4:1015 (2012)商山; J Biomed Res. 30: 2村窗09 (2016) | View |
| mlgG-Fc | The constant region (C年藍H2 and CH3 domains) of mouse immunog飛道lobulin heavy chain | Increases protein yie請都ld; allows rapid an吧請d simple detection of protein ex男草pression; enables simple 遠很protein G affinity chromato他請graphy based purification o線兵f Fc-tagged proteins. | EMBO Mol Med. 4:1015 (2012); Sci Rep. 6:30171 (2016)見睡 | View |
| NpuC(HN) | A modified C-terminal segm議區ent of the Nostoc punc房明tiforme (Npu) DnaE split intein | Facilitates purification of N-te森動rminal tagged proteins by in來秒tein-mediated cleavage and subsequent r火內elease of tagless ta明短rget proteins induced by a simple pH sh樂水ift (8.5 to 6.2) which leads to a cy筆上clization reaction of the C-terminal 下中asparagine to f刀還orm a succinimide ring. | Curr Protoc Protein Sci動報. 91:5.29.1 (2018) | View |
蛋白酶切割位點
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| EK | Bovine enterokinase cleavage site | Cleaved by enterokinase, which女就 cuts after lysine 數議in the recognition 南動sequence DDDDK. | Appl Microbiol Bio玩拍technol. 60:523 (2003) | View |
| TEV | Tobacco etch virus protease clea間拿vage site | Cleaved by toba文件cco etch virus protease, which cuts aft音日er glutamine in the recognit村文ion sequence EXXYXQ(G/S) where X can be風去 any amino acid. | Appl Microbiol 購女Biotechnol. 60:523 (2003) | View |
| Thrombin | Thrombin cleavage site | Cleaved by thrombin, which cu志那ts after the fourth re東討sidue in the recognition seque喝間nce XXP(R/K)XX where X通用X on the N end are hydrop腦場hobic residues and XX o哥近n the C end are 從裡non-acidic residues. | Appl Microbiol Biote動喝chnol. 60:523 (2003) | View |
| Factor-Xa | Factor Xa cleavage年會 site | Cleaved by factor Xa, which cuts after 很拿arginine in the recognition sequence I(國市E/D)GR. | Appl Microbiol Biotechnol. 60:523 (歌費2003) | View |
| HRV3C | Human rhinovirus 3C protease sit作風e | Cleaved by human rhinovir件車us 3C protease, the cleavage site i工校s between Gln and Gly i費近n the recognition sequences 煙說LEVLFQ↓GP. | Cell.168(3):427-441 (2017) | View |
信号肽 (for directing subcellular頻也 localization of proteins, such 白吧as nuclear localization, memb錢們rane localization, s醫光ecretion, etc.)
| 姓名 | 描述 | Application Note火新s | References | 序列 |
|---|---|---|---|---|
| SV40-NLS | Simian virus 40謝美 nuclear localizat厭影ion signal | Directs nuclear localization of tagged 明空protein. | Cell. 39:499 (1事聽984) | View |
| 3xNLS | 3 tandem simian 飛物virus 40 nuclear localiza可近tion signal | Directs nuclear localiz媽秒ation of tagged protein. | J Immunol. 182: 3173 (2009一分) | View |
| cMYC-NLS | Nuclear localization sign草要al of human c-MYC gene | Directs nuclear localization of tag腦劇ged protein. | Mol Cell Biol. 8:4048 (1988);Bioconjug Chem. 26: 1004 (20林不15) | View |
| BPSV40-NLS | A classical nuclear loc時跳alization signal (NLS) consists of t麗水wo clusters of basic res嗎員idues separated by 10–12 residues 空快(bipartite), NLS-conta明師ining cargoes are imported by a het樹科erodimeric import receptor 動樂complex composed of白我 importin α and importi快低n β | Directs nuclear lo地山calization of tagged protein. | J Biol Chem. 276:1317 (2001); Biophys J. 96:384購們0 (2009); Nature. 540:144 (2016); Nat Methods. 16:887-893 (這紙2019) | View |
| MITO | Mitochondrial ta書紅rgeting sequence fro書熱m human cytochrome c oxidase subunit VI答新II gene | Directs tagged p裡不rotein to the intermembran拿去e space of mitochond山白ria. | Nature. 358:325 (1992)森藍; Proc Natl Acad Sci U S A關北. 96: 11752 (1999) | View |
| Igκ-sig | Murine Ig kappa chain leader sequence | Directs tagged 年外protein to the secretory pathway. | J Immunol Methods. 152:89 (1森見992) | View |
| α-Factor | Yeast pheromone α窗山-factor leader sequence | Directs export of tagged道在 protein in yeast into cultu做都re medium. | Gene. 54:175 (1987) | View |
| HBM | Honeybee melittin signal peptid好頻e | Allows enhanced secretion of 票行tagged proteins from insect cells這票 into the mediu用機m. | Gene. 98:177 (1991) | View |
| PelB | Pectate lyase B leader 購土sequence | Directs tagged protein to 小費the bacterial periplasm, where the sequ的刀ence is removed by a s行通ignal peptidase. | Protein Expr Purif. 54:227 (2007) | View |
| NES | Nuclear export sign風草al | Directs nuclear export of 購那tagged protein. | PLoS One.10:e0137652 (2015) | View |
| Lyn | N-terminal myristoyla小農tion and palmitoylation seque白她nce from Lyn, a Src fa能匠mily tyrosine kinase | Anchors the tagged 綠開protein to the plasma membrane. | J Neurosci Methods. 93:37 (1999)媽地; J Cell Biol. 174:255 (2006) | View |
| SNAP25 | A protein which is synthesize農器d on free ribosome這湖s and recruited to the inner surface做個 of plasma membrane after the palimi快問toylation of specific cy話家stesine residues | Directs the tagged proteins to t些北he inner surface of plasma membrane. | EMBO J. 16:1575 (1997)電商 | View |
| GPI | Plasma membrane glycosy不工lphosphatidylinosito空歌l anchoring signa哥相l | Anchors glycoprotein to the plasm資朋a membrane; helps increase the mobilit冷票y of membrane proteins the的愛refore supporting signal tran體為sduction and cellu子又lar transport; play身風s an important role吃湖 in creating ant去地igens on cell surface. | Genesis. 44:202 (2006) | View |
| TMD-ST | Transmembrane domai這秒n of rat liver sialyltrans訊自ferase | Directs the tagged protei文玩n to Golgi. | J Biol Chem. 270:5483 (1995)讀爸 | View |
| PDGFR | Platelet-derived growth factor receptor站費 beta transmembrane domain | Anchors the tagged請人 protein to the extrac空玩ellular side of plasma membrane for dis家快playing, usually works with s小兒ecretory signal peptide機話. | Proc Natl Acad S對又ci U S A. 85:3435 遠機(1988); J Biol Chem. 285:12268 (2金遠010) | View |
| GLuc | Gaussia luciferase secretory signal p姐唱eptide | Allows the tagged protein to be好近 secreted from cytos動到ol. | Microb Cell Fact. 14:費火20 (2015) | View |
| IL2-sig | Signal peptide of 聽朋Homo sapiens interleukin議北 2 | Facilitates secretion of protei頻木ns. | Protein Expr Purif.118:105-12(2016) | View |
| HA-sig | The cleavable signal做開 sequence of influenza hemagglu路地tinin | Enhances locali廠討zation of proteins to the endoplasm白相ic reticulum memb對低rane or plasma membr紙讀ane. | J Biol Chem. 267:21995 (1992); Biochem Biophys Res 老哥Commun. 409:544 (2011) | View |
檢測标簽
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| TC | Tetracysteine tag | Facilitates detection謝紙 of tagged protein by biars計一enical compounds 數要such as FlAsH-EDT2 and ReAsH要關-EDT2, which turn fluorescent對亮 upon binding to the 文中tag. | Nat Protoc. 5: 1666 (高購2010) | View |
| ProS2 | Protein S of Myxococcus xanthu少兒s | Facilitates detection of tagged protei呢遠n by anti-ProS2 antibody; i跳了ncreases the solubility and stab謝妹ility of tagged pro術放tein in bacteria.雜中 | J Mol Biol. 286:1533 (1999) | View |
| TF | E.coli trigger factor, a ribosome-a金報ssociated chapero說人ne | Assists protein呢要 folding; facilitat金站es detection of tagged protein by鐵看 anti-trigger facto在這r antibody; increases聽我 the solubility of tagged protein. | Biochim Biophys A黑弟cta. 1803:650 (201請技0) | View |
| Lifeact | A 17-aa F-actin-binding sequence deriv答麗ed from Saccharomyc黑店es cerevisiae | Allows visualization of F-actin資聽 dynamics in cell媽暗s; can be used as a universal marker月冷 for F-actin imaging or as a那地n alternative to phalloidin. | Nat Methods. 5:605 (2008) | View |
| SNAP | A 20 kDa engineered variant o公到f the human DNA repair 朋微protein O-6-alky數歌lguanine-DNA alkyl吧火transferase, optimi笑有zed for mammalian expression | Allows to observe local錯少ization and transloca做也tion of SNAP-tag fusion遠懂 proteins; reacts specifically and麗到 rapidly with benzy分制lguanine (BG) and benzylchloro說拿pyrimidine (CP) derivatives, leading to如對 irreversible covalent labe花人ling of the SNAP-tag with a wide variet場司y of synthetic probes in坐謝 live cells. CLIP and SNAP fusion prot上請eins can be simultaneously and路紙 specifically labele上人d with different syntheti朋風c probes within cells. | Nat Biotechnol. 21:86 (2003); Methods Mol Biol. 也報1266:107 (2015) | View |
| CLIP | An engineered variant of the hu校話man DNA repair p多校rotein O-6-alkylguanine-DNA alkyltransf玩知erase, optimized for mammalian expr城雜ession | Allows to observe localization and t信能ranslocation of CLIP-tag fusi為暗on proteins; reacts speci吃月fically and rapidly with O2-b廠鐘enzylcytosine (著畫BC) derivatives, leading to irrever分樹sible covalent labelin近費g of the CLIP-tag wit笑湖h a wide variety of syntheti湖都c probes in live cel影市ls. CLIP and SNAP fusion prot民紅eins can be simultaneously and spec服妹ifically labeled with different輛湖 synthetic probes車器 within cells. | Chem Biol. 15:128 (2008) | View |
| ACP | Acyl carrier protein of E. coli | Allows to observe 為器localization and tran男街slocation of ACP-tag fusi東快on cell surface proteins which are spec好志ifically labeled with CoA de也些rivatives through post-translation很北al modifications catalyzed他行 by the phosphopantet謝煙heinyl transferase Ac科雪pS. | J Am Chem Soc. 126:8896 家麗(2004) | View |
| HiBiT | A small 11 amino acid peptide th筆計at binds LgBiT wi有懂th high affinity | Facilitates detection and monitoring of笑媽 tagged endogenous protein in live cel們新ls; capable of producing highly active 還看Nanoluc luciferase through high affi姐她nity complementation with LgBiT.姐道 | Biochem Biophys 土暗Rep. 12: 40 (2017); Chem Biol. 13:467 (2018) | View |
降解決定子标簽
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| DD | Destabilizing doma空電in, mutant derived from human子相 FKBP12 protein.靜習 | Targets the tagged protein for pr頻笑oteosomal degradation; addition of a c還房ompound, Shield-1, can stabilize the 間服protein and prevents費道 degradation. | Bioorg Med Chem Le物時tt. 18:5941 (2008); PLoS One. 4: e6529 (2音訊009) | View |
| S4D | Sal-like protein 4 (SALL4) d南我egron (S4D) tag | Allows degradation of S4D-tagged黃拍 proteins in the presence of immuno頻著modulatory drugs (IMi器討Ds) which interact with cereblon (很有CRBN) in the cullin E3 ubiq又紅uitin ligase complex to i開睡nduce degradation of neosubstrate南就s. | Commun Biol. 3:515 (2020) | View |
二聚體結構域
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| FKBP(F36V) | FK506-binding protein冷議 12 with F36V mutation | Promotes dimeriza歌白tion of tagged proteins in事店 the presence of dimerizer ligands照購, e.g. AP20187. | Chem Rev. 110:3315科這 (2010); Sci Rep. 4: 6127 (2014) | View |
細胞穿透肽
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| TAT | Amino acids 48–60 of transactivator o吧們f transcription protein (Tat靜生) from human immunodeficie妹身ncy virus type 1 (開開HIV-1). | Directs tagged protein to pass throu師看gh the plasma membrane of mammalian學會 cells. | J Biol Chem. 276:5836 (2001); ACS Nano. 8:197內歌2 (2014); Biomol Concepts. 8:131 (201房報7) | View |
| R8 | Octaarginine tag (8 tandem argenines).嗎生 | Directs tagged protein 玩去to pass through the plasm討就a membrane of mammal從靜ian cells. | J Biol Chem. 276:5836書山 (2001); ACS Nano. 8:1972 (2014慢光); Biomol Concepts. 8:131 (2017) | View |
轉錄調控蛋白
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| VP16 | VP16 transcriptional activator do理黃main encoded by HSV | Activates gene expression, commonly use土理d with other DNA binding proteins體件. | Cell. 149:1447 (2012) | View |
| VP64 | Tetrameric repeat of t那懂he minimal activa計件tion domain of VP16樹跳 | Strongly activates gene expr們商ession, commonly used 妹妹with other DNA bindi書船ng proteins. | Proc Natl Acad Sci U 地窗S A. 95: 14628 (1998); Proc Natl Acad Sci U S A店喝. 97: 1495 (2000) | View |
| KRAB | Krüppel-associated box d事到omain from human gen朋森e ZNF10 | Inhibits gene expression, commonly u子河sed with other DNA binding區土 proteins. | Proc Natl Acad Sci U S A. 91:4509 (快北1994); Proc Natl Acad S睡站ci U S A. 97: 1495 (2000) | View |
嵌合抗原受體結構域
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| CD8-leader | Leader signal peptide of T-cel員笑l surface glycopro雪拿tein CD8 alpha ch在議ain | Directs transport and local答又ization of the protein to the T-ce農金ll surface. | Cell. 64:891 (1991)哥道; WO2014055771A1; US20160185862A1; US10144770B2 | View |
| CD8-hinge | Hinge region of 你你T-cell surface gly小放coprotein CD8 alpha chain | Connects scFv and the t月自ransmembrane region; enhances t友得umor recognition efficiency; improves t雨業he expansion of CAR-T cells. | Cell. 64:891 (1991)湖資; J Immunother. 28:203 (2005); BMJ Open. 6: e013904 (2016); J Hematol Oncol. 10: 68 (2017); Oncoimmunology. 7: e13錯熱68604 (2018) | View |
| CD28-hinge | Hinge region of the T-cell surface 做件glycoprotein CD28 | Connects scFv with the t畫聽ransmembrane region providing stabilit議麗y and flexibilty for eff路技icient CAR expr和務ession and func廠煙tion; enhances efficiency of tumor 知公recognition; improves e吃房xpansion of CAR-T ce玩快lls. | J Immunother. 32:689 (200見些9); Mol Ther. 25:245不錢2 (2017) | View |
| CD8-TM | Transmembrane region of T-雨時cell surface glyco也雨protein CD8 alpha cha船內in | Anchors the CAR to the plasma membra年見ne, bridges the extracellular hinge生紙 and antigen recognition domain黑就s with the intrace票家llular signaling region化到. | Cell. 64:891 (1991); BMJ Open. 6: e013904 (2016); Oncoimmunology. 7: e1368604 (2018); WO2014055771A1; US20100105136A1 | View |
| CD28-TM | Transmembrane region歌紅 of T-cell-surface glycoprotein CD28 | Anchors the CAR to the 醫門plasma membrane; enhances re相門ceptor expression a司子nd stability. | Hum Gene Ther. 24:259 (2013); Cancer Res. 74:6383 (2自光014); WO2014055771A1; US10144770B2 | View |