首页
我的蚂蚁淘
快速下单
产品分类
- 分子类
- 蛋白类
- 抗体类
- 试剂盒
- 细胞相关
- 生化试剂
- 仪器类
- 诊断原料
- 其它
网站导航
- 特色主题 -
  
  秒杀抢购新品试用购物有礼精品推荐蚁豆商城品质团购活动专题 "1"元拍卖
- 蚂蚁淘 +
  
  体验官万众创业品牌合作我在蚂蚁淘
- 行业信息
  
  新闻资讯蚂蚁淘知道技术文章
- 客户服务
  
  帮助中心在线客服购物指南找货助手
关注我们
联系我们

主营：体外转录、基因克隆（BAC克隆试剂盒）、感受态细胞（TG1噬菌体文库构建感受态）、体外转录、转座子突变、蛋白表达

当前位置：首页 > 产品中心 > Bacterial Expression > Lucigen/Expresso® T7 Cloning and Expression System, N-His/49001-2/10 rxns

公司介绍

美国Lucigen公司，自1998年成立至今，一直致力于生命科学领域相关科研产品的研究与开发，在分子生物学领域处于领导性地位。Lucigen公司主要开发各类用于基因克隆的试剂盒及相关产品，包括：CloneSmart®平端克隆试剂盒、BigEasy®线性克隆系统、pEZSeq™平端克隆试剂盒、ClonePlex™ AK文库构建试剂盒、DNA Terminator ®末端修复试剂盒、EconoTaq® DNA聚合酶及E.cloni感受态细胞等�

产品分类

更多…

销售排行

更多…

问答列表

更多…

暂无问答

10 rxns

产品编号： 49001-2
美元价： $382.00
市场价： ¥7640.00
品牌： Lucigen
产地：美国(厂家直采)
公司： Lucigen
产品分类：转录表达>克隆和表达>分子类>
公司分类： Bacterial Expression

联系 Q Q: 3392242852
电话号码: 4000-520-616
邮箱: info@ebiomall.com

Lucigen/Expresso® T7 Cloning and Expression System, N-His/49001-2/10 rxns

商品介绍

CloneandexpressevendifficultproteinswiththefastestandeasiestdirectionalPCR-basedcloningsystemavailable.

Savetime!Enzyme-freecloninginsecondswithcloning-readyvectorandcells.
Highefficiency:›90%recombinants.
Tightly-controlledexpressionofN-orC-terminal6xHistaggedproteins.

AlsoavailablewithcleavableSUMOSolubilityTag

CompleteCloningandExpressionSystem

TheExpressoT7CloningandProteinExpressionSystemsaredesignedforfast,easy,andefficientdirectionalcloningandexpressionofPCR-amplifiedgenes.Thesystemsarecompletewithpre-processedpETite™N-orC-Hiscloningvectors,andtwocompetentcelllines,suppliedinsingletransformationvials.HighefficiencyHI-Control™10GChemicallyCompetentCellsenablestablecloningandHI-ControlBL21(DE3)CompetentCellsprovidetightlycontrolledproteinexpression,thushelpingyouavoidexpressionproblemsseenwithleakyT7promoter-basedsystems.TheN-orC-terminal6xHistaggedproteinscanberapidlypurifiedusingstandardNichromatography.SystemsarealsoavailablethatencodeanN-terminal6xHisSUMOtagfusionforimprovedsolubilityofexpressedprotein.OtherExpressoSystemsareavailablewhichexpressproteinsunderthecontroloftheRhamnosepromoter.

Five-SecondDirectionalCloningofPCR-AmplifiedGenes

TheExpressoT7CloningandProteinExpressionSystemusesExpressioneeringTechnology™,anenzyme-freerecombinationalcloningstrategytoseamlesslyintegratethegenewiththevector.ThetargetgeneisamplifiedbyPCRusingprimersthatadd18base-pairsofvector-complementarysequencetobothendsofthegene.Unlikeotherrestrictionenzymebasedmethodsorligase-freecloningmethods,nofurthercleanuporenzymatictreatmentofthePCRproductisnecessary.Simplymix1µloftheunpurifiedPCRreactionwiththesuppliedpre-processedpETite™T7expressionvector,andtransformimmediatelyintotheHI-Control™10GChemicallyCompetentCellsprovided(Figure1).

Expresso saves you a day!

Cloning-ReadyVectorsSaveHoursofResearchTime.

NewpETiteT7vectorsinclude:

StrongT7promoterforhigh-levelexpression.
ChoiceofN-terminalorC-terminal6xHisfusiontagsforfastproteinpurification.
Smallsize(2.2kb)foreasierdownstreammanipulation.
PatentedCloneSmart®technologyincreasescloningefficiency.

pETite vector maps

Figure2.pETiteT7expressionvectors:Smallsize(2.2kbvs.5.4kbforpET)foreasiermanipulation,includingtargetedmutagenesis.Vectorsarepre-processedforinstantenzyme-freecloningofPCRproductswithachoiceofamino-terminalorcarboxyl-terminalfusionof6xHistagtoproteinofinterest.

HighcloningefficiencywithHI-Control10GChemicallyCompetentCells.

ThehightransformationefficiencyofHI-Control10GChemicallyCompetentCellsensuresrecoveryofcloneswithprecisejunctionsandthecorrectorientation.Formostgenes,>90%ofcolonieswillhavethetargetgeneinsertedinthecorrectorientation.(Figure3).

Figure3.HighcloningefficiencywithExpressoT7System.
Pre-processedpETiteC-Hisvectorwasmixedwith1µlofunpurifiedPCRgeneproductandtransformedintoHI-Control10GChemicallyCompetentCells.ColonyPCRwasperformedon18randomlychosencolonies;17of18containedinsertofthecorrectsize.

HI-ControlBL21(DE3)CellsControlLeakyProteinExpression

HI-ControlBL21(DE3)CellscontainhighlevelsoflacrepressortomaintaintightcontroloverexpressionofT7RNApolymerase.Tightercontrolmeansbettertoleranceofpotentiallytoxicgeneproducts(Figure4).

Figure4.ExpressionofvariousproteinsusingtheExpressoT7SystemversusapETVector.GenesencodingaDNApolymerase,abluefluorescentprotein(BFP),orATPsynthasebsubunit(membraneprotein)wereclonedintopET28aorpETitevectorswithN-terminalorC-terminal6xHistags(asindicated).pET28aclonesweretransformedintostandardBL21(DE3)cells,andpETiteclonesweretransformedintoHI-ControlBL21(DE3)cellsforexpression.CulturesweregrowninLBat37°CtoanOD₆₀₀of0.5to0.7(odd-numberedlanes)andinducedfor3hourswith1mMIPTG(even-numberedlanes).CellswerepelletedandlyseddirectlyinSDS-PAGEloADIngbuffer,and0.05ODequivalentswereanalyzedbygelelectrophoresis.ThegelwasstainedwithCoomassieblue.

PurificationofActiveSolubleFluorescentProteinUsing6xHisTag

TheN-orC-terminal6xHistaggedproteinsexpressedusingtheExpressoT7CloningandExpressionSystemcanberapidlyaffinity-purifiedovercommerciallyavailablenickelresins.Anexampleofthepurificationofa6xHistaggedyellowfluorescentproteinclonedandexpressedusingtheExpressoT7SystemisshowninFigure5.

Expression and purification of active soluble fluorescent protein.

Figure5.Purificationofa6xHistaggedfluorescentprotein.HI-ControlBL21(DE3)cellsharboringpETiteC-Hisvectorcontainingayellowfluorescentprotein(YFP)geneweregrownat37°CinLBmediatoanOD₆₀₀of0.6(lane1),theninducedwith
1mMIPTGfor4hours(lane2). Cellswereharvestedandlysedbysonicationin300mMNaCl,50mMTris-HClpH8.0.ClearedlysatewasloadedontoanNi-NTASepharose®Column.Columnflow-through(lane3,FT)andwash(lane4,W)fractionswerecollected.TheboundYFP,showninthecolumnatleft,waselutedwithwashbuffercontaining300mMimidazole(lanes5-12,E1-E8).Fluorescentfractionsalignwiththepureproteinasshownonthegel.

ImportantProductUseInformation:
ThisproductisthesubjectofU.S.Patent#6,709,861.AdditionalpatentapplicationsownedbyLucigenCorporationarepending.

The6xHistagislicensedfromHoffmann-LaRoche,Inc.,Nutley,NJand/orHoffmann-LaRocheLtd.,Basel,Switzerlandandisprovidedonlyfortheuseinresearch. InformationaboutlicensesforcommercialuseisavailablefromQiagenGmbH,QIAGENStrasse1,D-40724Hilden,Germany.Purificationofthe6xHistaggedproteinswithNi-NTAmanufacturedbyQIAGENishighlyrecommendedforbestperformancesandtoavoidpoorpurificationresults.

ORDERINFORMATION

TheExpressoT7Cloning&ExpressionSystemcontainspre-processedpETite®N-Hisand/orpETiteC-HisVectorDNA,HI-Control™10GChemicallyCompetentCellsforcloning,andHI-ControlBL21(DE3)ChemicallyCompetentCellsforproteinexpression.AlsoincludedareN-Hisand/orC-HisPositiveControlInsertDNAs,andtransformationpositivecontrolpUCDNA.

TheExpressoT7SUMOCloningandExpressionSystemcontainspre-processedpETite®N-HisSUMOVectorDNA,HI-Control™10GChemicallyCompetentCellsforcloning,andHIControlBL21(DE3)ChemicallyCompetentCellsforproteinexpression.AlsoincludedareSUMOPositiveControlCInsertDNA,transformationpositivecontrolpUCDNA,SUMOExpressProtease,SUMOCleavageControlProtein,andforwardandreversePCRprimerstoconfirmclones.

品牌介绍

蚂蚁淘(ebiomall）作为一家生命科学领域的垂直电商平台，公司专注生物医学科研用品的全球导购和品牌推广，为海内外厂方与代理经销商搭建跨境贸易系统和交流平台。产品齐全，200+分类，1000+品牌，500万+产品信息。100%正品，专业售后，到货快，在线下单，简单轻松，节省科学家宝贵的科研时间。为您竭诚服务！

【订购】蚂蚁淘生物科技联系方式：4000-520-616

更多产品，更多优惠！请联系我们！

苏州蚂蚁淘生物科技有限公司

免费热线：4000-520-616

官网：https://www.ebiomall.com/

Lucigen有各种各样的克隆系统，包括新型的无转录克隆载体，可提高克隆产量，降低克隆偏倚并提高测序通量。为了克隆最困难的序列，BigEasy®v2.0线性克隆系统提供了无与伦比的稳定性。大多数Lucigen克隆载体都是预先切割，去磷酸化的，可以立即使用。