| Discount | $100DiscountCodeforYourFirstOrderof1mgCreRecombinase:CreDiscount Discountforswitchingfromcompetitors'product:$150.EmailtoTechSupportts@Excellgen.comfordiscountcode. |
| Description | Crerecombinase,oftenabbreviatedtoCre,isaTypeItopoisomerasefromP1bacteriophagethatcatalyzessite-specificrecombinationofDNAbetweenloxPsites.Thisenzymedoesnotrequireanyenergycofactors,andCre-mediatedrecombinationquicklyreachesequilibriumbetweensubstrateandreactionproducts.TheloxPrecognitionelementisa34basepair(bp)sequencecomposedoftwo13bpinvertedrepeatsflankingan8bpspacerregionwhichconfersdirectionality.RecombinationproductsaredependentonthelocationandrelativeorientationoftheloxPsites.TwoDNAspeciescontainingsingleloxPsiteswillbefusedwhilstDNAbetweenloxPsitesinthesameorientationwillbeexcisedincircularformandDNAbetweenopposingloxPsiteswillbeinvertedwithrespecttotherestoftheDNA. Crerecombinaseisusedasatooltomodifygenesandchromosomes.InthisapproachtheCrerecombinaseisusedtodeleteasegmentofDNAflankedbyLoxPsites(aka'floxed')inanexperimentalanimal.Ithasbeenusedtogenerateanimalswithmutationslimitedtocertaincelltypes(tissue-specificknockout)oranimalswithmutationsthatcanbeactivatedbydrugadmiNISTration(inducIBLeknockout)inanumberoftransgenicspecies.TheavailABIlityoftransgeniclineswithtissuespecificorinducibleCreexpressionpermitsresearcherstoinactivateoractivateageneofinterestsimplybybreedingafloxedanimaltopre-existingCre-transgenics.OneexampleofaninducibleCrerecombinasesystemistheCre-ER(ER=EstrogenReceptor)systeminwhichintraperitonealinjectionoftamoxifenwillcausedose-dependentexcisionofthefloxedsite(i.e.willinactivatethegeneofchoice). RecombinantCrerecombinase(TAT-Cre)waspurifiedfromanE.colistraincarryinganengineeredplasmidencodingenhancedformofCreRecombinasefrombacteriophageP1.ThisCrerecombinasehasanN-terminal6XHistag,aTatpeptide(GRKKRRQRRRPPAGTSVSL)andanNLSsequence(PKKKRKV).HTNCisthemosteffectiveproteinintransduction(invivo)andsubsequentrecombinationcomparedtootherformsofCrerecombinases,e.g.,HNC,TCH6,HC,HNCM,CH.Incubationoffibroblastreportercellswith1μMHTNCfor1to2hourscanresultintranductionof60~90%ofthecells*.Additionof100μMchoroquinetoculturemediumcanfurtherenhancetransductionandrecombination. *Usethispagetocalculateconcentrationrequiredforculturedcells: http://www.excellgen.com/pub/mw_2_moles.html |
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| Properties | MolecularWeight:43kDa QC:Tat-Creinduced80~100%recombinationefficienciesinHEK293T-Crereportercells.HEK293T-cellsweretransfectedwithreporterplasmidDNA:LoxP-RFP-Stop-LoxP-GFP,thenadd1μMpurifiedTat-Creprotein.
Fig1:0:beforeaddingTat-Cre;1to5:1to5daysaftertransductionwithTat-Cre.
Fig2:ComparisonofrecombinationacitivitiesofTat-Crerecombinaseinliquidand lyophilizedform.1,Tat-Crerecombinaseinliquidformafterstoringat-80oCfor6months;2,LyophilizedTat-Crerecombinaseafterstoringat37oCfor2weeks;3,LyophilizedTat-Crerecombinaseafterstoringat37oCfor2weeksandsubsequentlystoringat4oCfor2weeks.Picturesweretaking15hoursafteradding1uMTat-Crerecombinase. EndotoxinLevels:<0.1EU/μg Purity:>98%bySDS-PAGEandHPLCanalysis. |
| UnitDefintion | Astandardof100UnitsisdefinedastheamountofTAT-CRE(μg) in1.0mLoftissueculturemediumthatisrequiredtoinduce50% GFPexpressioninaHEK293Treportercelllineassay. |
| Stability | 5yearsat-80oC.2yearsat-20oC.1yearat4oC,upto6monthsatambienttemperature |
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| Storage | 4~-20ºC |
| Shipping | ambienttemperature(reducedshippingfee) |
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尽量简洁
缺点：质粒仍然较大，转染难度相对较大。具有碱基识别偏好性，局限了基因编辑的运用范围，而且会导致不同基因位点编辑效率不同。筛选仍然需要较大工作量。
http://www.nature.com/nature/journal/v520/n7546/full/nature14299.html
1、过表达目的蛋白，用以检测基因表型；
2、荧光标签标记目的蛋白，用以跟踪其细胞 定位；
3、引入野生型等位基因。
是的，确切来说是大量表达。 大肠杆菌是基因重组技术中常用的细菌，将外源目的基因(如人胰岛素基因)导入大肠杆菌后可在大肠杆菌内表达目的蛋白(如胰岛素)，由于细菌繁殖速度快，通过发酵便可在短时间内获得大量胰岛素，再经多步分离、纯化便得到了药用胰岛素。
流程大概是这样的：首先获得小鼠ES细胞系，测试ES细胞嵌合入受体囊胚的能力之后根据不同基因、不同目的设计并构建打靶载体，将打靶载体转入一定数目ES细胞中，然后鉴定出带有发生正确同源重组的突变中靶ES细胞。通过显微注射或者胚胎融合的方法将经过遗传修饰的ES细胞引入受体胚胎内。经过遗传修饰的ES细胞可以发育为嵌合体动物的生殖细胞，是的经过修饰的遗传信息经生殖系遗传，从而得到带有修饰基因的突变小鼠，而后可以对其进行表型分析。
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