常用pGEX载体图谱

Rosetta系列的表达菌株可以提供T7RNA聚合酶，它能表达PET系列载体上的外源基因。。。pGEX系列载体上的外源基因不需要T7RNA聚合酶，普通的大肠杆菌经IPTG诱导即可表达Tac启动子是一组由Lac和trp启动子人工构建的杂合启动子，受Lac阻遏蛋白的负调节，它的启动能力比Lac和trp都强。其中Tac1是由Trp启动子的－35区加上一个合成的46bpDNA片段(包括Pribnow盒)和Lac操纵基因构成，Tac12是由Trp的启动子-35区和Lac启动子的-10区，加上Lac操纵子中的操纵基因部分和SD序列融合而成蛋白标签：Amyctagisapolypeptideproteintagderivedfromthec-mycgeneproductthatcanbeaddedtoaproteinusingrecombinantDNAtechnology.Itcanbeusedforaffinitychromatography,thenusedtoseparaterecombinant,overexpressedproteinfromwildtypeproteinexpressedbythehostorganism.Itcanalsobeusedintheisolationofproteincomplexeswithmultiplesubunits.Amyctagcanbeusedinmanydifferentassaysthatrequirerecognitionbyanantibody.Ifthereisnoantibodyagainstthestudiedprotein,addingamyc-tagallowsonetofollowtheproteinwithanantibodyagainsttheMycepitope.ExamplesarecellularlocalizationstudiesbyimmunofluorescenceordetectionbyWesternblotting.Thepeptidesequenceofthemyc-tagis(in1-and3-lettercodes,respectively):N-EQKLISEEDL-C,N-Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-C,whereNstandsforAmino-terminusandCstandsforCarboxyterminus.Thetagisapproximately1202Daltonsinatomicmassandhas10aminoacids.ItcanbefusedtotheC-terminusandtheN-terminusofaprotein.Itisadvisablenottofusethetagdirectlybehindthesignalpeptideofasecretoryprotein,sinceitcaninterferewithtranslocationintothesecretorypathway.Amonoclonalantibodyagainstthemycepitope,named9E10,isavailablefromthenon-commercialDevelopmentalStudiesHybridomaBankpGEX4T1载体基本信息出品公司:GE别名:pGEX-4T-1,pGEX4T1,pGEX4T1质粒类型:大肠杆菌蛋白表达载体表达水平:高拷贝启动子:Tac克隆方法:多克隆位点，限制性内切酶载体大小:4969bp5'测序引物及序列:pGEX5':GGGCTGGCAAGCCACGTTTGGTG3'测序引物及序列:pGEX3':CCGGGAGCTGCATGTGTCAGAGG载体标签:N-GST载体抗性:Ampicillin氨苄备注:复制子是pMB1产品目录号:27-4580-01稳定性:瞬时表达Transient组成型:诱导表达病毒/非病毒:非病毒pGEX4T1载体质粒图谱和多克隆位点信息原核生物DNA复制起始点，是DNA链上独特的具有起始DNA复制功能的碱基序列。大肠杆菌的复制起点包括OriC和OriHThrombin凝血酶pGEX4T1载体简介pGEX4T1载体序列LOCUSpGEX-4T-14969bpDNAcircularSYNDEFINITIONpGEX-4T-1ACCESSIONKEYWORDSSOURCEORGANISMothersequences;artificialsequences;vectors.COMMENTThisfileiscreatedbyVectorNTI|biofeng.com|FEATURESLocation/Qualifierssource1..4969/organism=pGEX-4T-1/mol_type=otherDNApromoter184..212/label=tac_promotermisc_feature224..246/label=M13_pUC_rev_primergene258..977/label=GST(variant)/gene=GST(variant)CDS258..977/label=ORFframe3misc_featurecomplement(1019..1041)/label=pGEX_3_primerpromoter1307..1335/label=AmpR_promotergene1377..2237/label=Ampicillin/gene=AmpicillinCDS1377..2237/label=ORFframe3rep_origin2392..3011/label=pBR322_originmisc_feature3309..4400/label=lacICDS3441..4400/label=ORFframe3promoter4449..4478/label=lac_promotermisc_feature4492..4514/label=M13_pUC_rev_primerpromoter4513..4531/label=M13_reverse_primerCDS4523..81/label=ORFframe2misc_feature4540..4695/label=lacZ_apromotercomplement(4543..4559)/label=M13_forward20_primermisc_featurecomplement(4552..4574)/label=M13_pUC_fwd_primerpGEX4T2载体基本信息出品公司:GE别名:pGEX-4T-2,pGEX4T2,pGEX4T2质粒类型:大肠杆菌蛋白表达载体表达水平:高拷贝启动子:Tac克隆方法:多克隆位点，限制性内切酶载体大小:4970bp5'测序引物及序列:pGEX5':GGGCTGGCAAGCCACGTTTGGTG3'测序引物及序列:pGEX3':CCGGGAGCTGCATGTGTCAGAGG载体标签:N-GST载体抗性:Ampicillin氨苄备注:复制子是pMB1产品目录号:27-4581-01稳定性:瞬时表达Transient组成型:诱导表达病毒/非病毒:非病毒pGEX4T2载体质粒图谱和多克隆位点信息pGEX4T2载体简介pGEX4T2载体序列LOCUSpGEX-4T-24970bpDNAcircularSYNDEFINITIONpGEX-4T-2ACCESSIONKEYWORDSSOURCEORGANISMothersequences;artificialsequences;vectors.COMMENTThisfileiscreatedbyVectorNTI|biofeng.com|FEATURESLocation/Qualifierssource1..4970/organism=pGEX-4T-2/mol_type=otherDNApromoter184..212/label=tac_promotermisc_feature224..246/label=M13_pUC_rev_primergene258..978/label=GST(variant)/gene=GST(variant)CDS258..974/label=ORFframe3misc_featurecomplement(1020..1042)/label=pGEX_3_primerpromoter1308..1336/label=AmpR_promotergene1378..2238/label=Ampicillin/gene=AmpicillinCDS1378..2238/label=ORFframe1rep_origin2393..3012/label=pBR322_originmisc_feature3310..4401/label=lacICDS3442..4401/label=ORFframe1promoter4450..4479/label=lac_promotermisc_feature4493..4515/label=M13_pUC_rev_primerpromoter4514..4532/label=M13_reverse_primerCDS4524..81/label=ORFframe3misc_feature4541..4696/label=lacZ_apromotercomplement(4544..4560)/label=M13_forward20_primermisc_featurecomplement(4553..4575)/label=M13_pUC_fwd_primerpGEX4T3载体质粒图谱和多克隆位点信息pGEX4T3载体简介pGEX4T3载体序列LOCUSpGEX-4T-34968bpDNAcircularSYNDEFINITIONpGEX-4T-3ACCESSIONKEYWORDSSOURCEORGANISMothersequences;artificialsequences;vectors.COMMENTThisfileiscreatedbyVectorNTI|biofeng.com|FEATURESLocation/Qualifierssource1..4968/organism=pGEX-4T-3/mol_type=otherDNApromoter184..212/label=tac_promotermisc_feature224..246/label=M13_pUC_rev_primergene258..976/label=GST(variant)/gene=GST(variant)CDS258..980/label=ORFframe3misc_featurecomplement(1018..1040)/label=pGEX_3_primerpromoter1306..1334/label=AmpR_promotergene1376..2236/label=Ampicillin/gene=AmpicillinCDS1376..2236/label=ORFframe2rep_origin2391..3010/label=pBR322_originmisc_feature3308..4399/label=lacICDS3440..4399/label=ORFframe2promoter4448..4477/label=lac_promotermisc_feature4491..45