质粒pECFP-N1图谱
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(转载)一.九种表达载体Pllp-OmpA,pllp-STII,pMBP-P,pMBP-C,pET-GST,pET-Trx,pET-His,pET-CKS,pET-DsbA 二.克隆载体pTZ19RDNApUC57DNAPMD18TPQE30pUC18pUC19pTrcHisApTrxFuspRSET-ApRSET-BpVAX1PBR322pbv220pBluescriptIIKS( )L4440pCAMBIA-1301pMAL-p2XpGD926三.PET系列表达载体ProteinExpression?ProkaryoticExpression?pETDsbFusio nSystems39band40bProteinExpression?ProkaryoticExpression?pETExpressi onSystem33bProteinExpression?ProkaryoticExpression?pETExpressi onSystemsProteinExpression?ProkaryoticExpression?pETExpressi onSystemsplusCompetentCellsProteinExpression?ProkaryoticExpression?pETGSTFusio nSystems41and42ProteinExpression?ProkaryoticExpression?pETNusAFusi onSystems43.1and44ProteinExpression?ProkaryoticExpression?pETVectorDN AProteinPurification?PurificationSystems?Strep?Tacti nResinsandPurificationKits四.PGEX系列表达载体TEcoR?pGEX-1I/BAP pGEX-2T pGEX-2TKpGEX-3XpGEX-4T-1pGEX-4T-2pGEX-4T-3pGEX-5X-1pGEX-5X-2pGEX-5X-3pGEX-6P-1pGEX-6P-2pGEX-6P-3五.PTYBsystemPTYB1PTYB2PTYB11PTYB12六.真核表达载体pCDNA3.1(-)pCDNA3.1( )pPICZalphaApGAPZαAPYES2.0pBI121pEGFP-N1pEGFP-C1pPIC9KpPIC3.5K如何阅读分析质粒图谱载体主要有病毒和非病毒两大类,其中质粒DNA是一种新的非病毒转基因载体。
(转载)欧阳歌谷(2021.02.01)一.九种表达载体Pllp-OmpA,pllp-STII,pMBP-P,pMBP-C,pET-GST,pET-Trx,pET-His,pET-CKS,pET-DsbA 二.克隆载体pTZ19RDNApUC57DNAPMD18TPQE30pUC18pUC19pTrcHisApTrxFuspRSET-ApRSET-BpVAX1PBR322pbv220pBluescriptIIKS( )L4440pCAMBIA-1301pMAL-p2XpGD926三.PET系列表达载体ProteinExpression?ProkaryoticExpression?pETDsbFusionSystems39b and40bProteinExpression?ProkaryoticExpression?pETExpressionSystem33b ProteinExpression?ProkaryoticExpression?pETExpressionSystems ProteinExpression?ProkaryoticExpression?pETExpressionSystemsplus CompetentCellsProteinExpression?ProkaryoticExpression?pETGSTFusionSystems41a nd42ProteinExpression?ProkaryoticExpression?pETNusAFusionSystems43 .1and44ProteinExpression?ProkaryoticExpression?pETVectorDNA ProteinPurification?PurificationSystems?Strep?TactinResinsandPurific ationKits四.PGEX系列表达载体TEcoR?pGEX-1I/BAP pGEX-2TpGEX-2TKpGEX-4T-1 pGEX-4T-2 pGEX-4T-3 pGEX-5X-1 pGEX-5X-2 pGEX-5X-3 pGEX-6P-1 pGEX-6P-2 pGEX-6P-3五.PTYBsystem PTYB1PTYB2PTYB11PTYB12六.真核表达载体pCDNA3.1(-) pCDNA3.1( ) pPICZalphaA pGAPZαA PYES2.0pBI121pEGFP-C1pPIC9KpPIC3.5K如何阅读分析质粒图谱载体主要有病毒和非病毒两大类,其中质粒DNA是一种新的非病毒转基因载体。
pAD-SV40T编号载体名称北京华越洋VECT76070pAD-SV40TpAD-SV40T载体图谱:pAD-SV40T载体简介:The pAD-SV40T control plasmid expresses a hybrid protein which contains the NF-κB transcription activation domain fused to amino acids84–708of the SV40large T-antigen.5pAD-SV40T载体相关的哺乳动物表达载体:SuperCos I pDsRed2-Bid pNFκB-MetLuc2-ReporterpYr-adshuttle-3pAcGFP1-N1pEF1α-IRES-DsRed-Express2pVitro2-neo-mcs pSecTag2A pCMV-DsRed-Express2pUB6/V5-His/LacZ pGL4.27pcDNA3.1/NT-GFP-TOPOpUB6/V5-His C pGL4.26pEF1α-IRES-ZsGreen1pUB6/V5-His B pACT pCMV-Tag2ApUB6/V5-His A pBIND-Id Control pCMV-Tag5BpTracer-CMV2pTRE2pAcGFP1-C In-Fusion ReadypSV-β-Galactosidase pRevTRE p3XFLAG-CMV-14pSI pTK-hyg p3XFLAG-CMV-8pSG5pTRE3G-Luc pFLAG-CMV-2pSFV1pSwitch pcDNA3.3-TOPOpSecTag2/Hygro A pcDNA4/His C pcDNA6.2/cLumio-DESTpSecTag B c-Flag pcDNA3pCMV-tdTomatopRluc-N2pcDNA4/TO/Myc-His A pAcGFP1-MitopPICZalpha D pcDNA6/myc-His B pAcGFP1-N In-Fusion Ready pORF-lacZ pcDNA6/V5-His B pDsRed-Monomer-N In-Fusion Ready pORF-HSV1tk pcDNA6.2/nTC-Tag-DEST pcDNA4/TO/Myc-His BpOG44pOptiVEC-TOPO pIRES2-EGFPpNTAP-B pcDNA5/FRT pcDNA3.1/His CpMEP4pGL4.30pcDNA3.1/CT-GFP-TOPOpLVX-ZsGreen-miRNA-Puro pGL4.19pEF1α-IRES-AcGFP1pLVX-IRES-Puro-3xFlag pACT-MyoD pcDNA3.2/V5/GW/D-TOPO pLPCX pCMV-BD pcDNA4/TO/Myc-His/LacZ pLEGFP-N1pCMV-Tet3G pcDNA4/HisMax-TOPOpKH3pTet on advanced p3XFLAG-CMV-13pIRES-puro2pTRE-Tight p3xFLAG-CMV-10phRL-TK pIND pFLAG-CMV-3pG5lac pGene/V5-His B pcDNA4/TO/Myc-His CpFR-luc pOPRSVI pcDNA6.2/C-YFP-DESTpEF6/myc-His C pcDNA4/HisMax A pcDNA6.2/cTC-Tag-DESTpEF4/V5-His A pIRESpuro3pcDNA6.2/nGeneBLAzer-DESTpEF1/myc-His lacZ pIRESneo3pCRE-MetLuc2-ReporterpEF1/myc-His C pIRESneo2pEF1α-DsRed-Express2pEF1/myc-His B pcDNA4/myc-His A pDsRed-Express-C1pEF1/myc-His A pCMV-PKA pEF1α-DsRed-Monomer-N1 pECFP-Mito pAcGFP1-N3pDD-AmCyan1ReporterpECFP-ER pcDNA5/FRT/TO pCRE-DD-AmCyan1pDP8rs pBApo-CMV-Pur pIRES2-DsRed-Express2pDP5rs pBApo-EF1α-pur pDsRed-Express-N1pDP4rs ptdTomato-N1pcDNA6.2/nLumio-DESTpDP3rs pAcGFP1-Golgi pcDNA6/myc-His ApDP2rs pAcGFP1-p53pcDNA6/V5-His ApDP1rs pAcGFP1-Actin pcDNA5/FRT/TO-TOPOpCMV-Myc-C pBI-CMV2pcDNA6.2/V5/GW/D-TOPOpCMV-HA-N pEF1α-tdTomato pcDNA6.2/cGeneBLAzer-DEST pCMV-HA-C pEF1α-tdTomato pcDNA6.2/nGeneBLAzer-GW/D-TOPO pCMV6-XL4pEBVHis A pcDNA5/TOpCMV6-AC-GFP pGL4.10pBApo-CMV-neopCMV5pGL4.29pDsRed-MonomerpCI pGL4.13pIRESpCGN pG5luciferase pIRES-hrGFP-1apcDNA6.2/EmGFP-Bsd/V5-DEST pCMV-AD pDsRed-Express2-N1pcDNA4/V5-His A pRevTet-Off pCMV-Tag3BpcDNA3-mRFP pTet-Off pCRE-hrGFPpcDNA3-CFP pTRE2-hygro pDsRED2-MitopcDNA3.1(+)/myc-His C pVgRxR pAcGFP1-FpcDNA3.1(+)/myc-His A pOPI3CAT pAcGFP1-C1pcDNA3.1(+)/CAT pBK-RSV pAsRed2-C1pcDNA3.1(-)/myc-His C pIRES2-DsRed2pAsRed2-N1pcDNA3.1/Zeo(+)pCMV-Myc pAcGFP1-LampcDNA3.1/Zeo(-)pCMV-Tag2C pAcGFP1-CpcDNA3.1/V5-His C pCMV-Tag5A pSEAP2-BasicpcDNA3.1/Hygro(+)pCMV-Tag3C pBI-CMV3pcDNA3.1/Hygro(-)p3XFLAG-CMV-7pNFkB-DD-tdTomatopcDNA3.1/GS p3XFLAG-CMV-9pcDNA3.1/His ApBS185CMV-Cre pFLAG-CMV-4pEBVHis BpBIND-GFP pBI-CMV4pGL4.75pBiFC-VN155(I152L)pcDNA4/His A pGL4.20pBiFC-CC155pcDNA4/myc-His B pCMV-SPORT6pBiFC-CrN173pcDNA4/HisMax C pCMV-SPORT6pBiFC-bJunVN155(I152L)pCMV-Tag4A pBINDpBD-p53pcDNA6/myc-His C pBD-NF-κBpBD-NF-kB pCMV-Tag2B pRevTet-OnpBC1pGRN145pTet-OnpAD-TRAF pCMV-MEK1pTRE3GpAD-SV40T pCMV-Tag3A pcDNA4/TOpAAV-ZsGreen-miRNA pCMVLacI pcDNA4/His B pAAV-tTS-shRNA pBI-CMV1pcDNA4/HisMax B pIREShyg3pEF1α-AcGFP1-N1pcDNA4/myc-His C pcDNA6/TR pCMV-LacZ pcDNA3.1/His B pDsRed-Express2-C1pCMV-Tag4B pcDNA6/V5-His C pBI-CMV5pCMV-Tag5C pCHO1.0 pAmCyan1-C1plRES2-ZsGreen1pGL3-Promoter pAcGFP1-Mem p3XFLAG-CMV-7.1pCMV-MEKK1 pAcGFP1-C3pFLAG-CMV-5a pFLAG-CMV2 pTT5pBudCE4.1pAcGFP1-C2 pSEAP2-Control pREP4ptdTomato-C1 pIRES2-AcGFP1pBApo-CMV pCRE-DD-tdTomato pAcGFP1-Hyg-C1。
)质粒图谱登记号:00012)质粒名称:pIRES3)来源:BD Co4)用途:真核双表达5)是否可以提供更详细资料:可以6)是否可以共享:7)联系方式:PM)质粒图谱登记号:00022)质粒名称:pECFP-C13)来源:BD Co4)用途:检测真核表达5)是否可以提供更详细资料:可以6)是否可以共享:7)联系方式:pm1)质粒图谱登记号:00032)质粒名称:pShuttle3)来源:4)用途:5)是否可以提供更详细资料:6)是否可以共享:7)联系方式:2)pShuttle MCS很多人用pEGFP-C1,我也来发一个)质粒图谱登记号:00042)质粒名称:pSBR322、pUC183)来源:4)用途:5)是否可以提供更详细资料:6)是否可以共享:7)联系方式:1)质粒图谱登记号:00052)质粒名称:pcDNA3.1(+)/CAT3)来源:invitrogen4)用途:真核表达5)是否可以提供更详细资料:可以6)是否可以共享:无偿7)联系方式:PM1)质粒图谱登记号:00062)质粒名称:pQEx3)来源:Qiagen4)用途:原核表达5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM2)1)质粒图谱登记号:00072)质粒名称:pIVEX2.33)来源:Rocho4)用途:体外转录翻译5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM质粒图谱登记号:0008质粒名称:pIRES-EGFP来源:用途:是否可以提供更详细资料:是否可以共享:否联系方式:1)质粒图谱登记号:00092)质粒名称:pET-28a(+)3)来源:Novagen4)用途:原核表达5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM1)质粒图谱登记号:00112)质粒名称:pET-32a(+)3)来源:novagen4)用途:原核表达5)是否可以提供更详细资料:6)是否可以共享:7)联系方式:pm1)质粒图谱登记号:00132)质粒名称:pcDNA3.1/Zeo (+)3)来源:invitrogen4)用途:原核表达5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM)质粒图谱登记号:00132)质粒名称:pEGFP-N33)来源:clontech4)用途:真核表达5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM)质粒图谱登记号:00142)质粒名称:pcDNA33)来源:invitrogen4)用途:真核表达5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM)质粒图谱登记号:00152)质粒名称:pfastbac13)来源:invitrogene4)用途:昆虫表达5)是否可以提供更详细资料:不可以6)是否可以共享:不可以7)联系方式:PM)质粒图谱登记号:00162)质粒名称:pEGFP-C33)来源:clontech4)用途:真核表达5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PMwangjun2002274 edited on 2004-06-22 00:041)质粒图谱登记号:00172)质粒名称:pSecTag23)来源:Invitrogen4)用途:真核表达5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM)1质粒图谱登记号:00192)质粒名称:pET20b3)来源:NOVAGEN4)用途:原核表达5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM1)质粒图谱登记号:00222)质粒名称:pThioHisA3)来源:invitrogen4)用途:原核表达5)是否可以提供更详细资料:4.365kb , HP-thioredoxin fusion proteinexpressionvector, trc promoter, Ampr, a EK cleavage site lies between HP-thioredoxin and MCS宿主菌TOP10(基因型为:F-mcrA △(mrr-hsd RMS-mcrBC)Ф80 lacZ M15 △lacX74 deoR recAl araD139 △(ara-leu)7697 galU galK rpsL endAl nupG6)是否可以共享:交换或其它7)联系方式:PM2)3)1)质粒图谱登记号:00242)质粒名称:pcDNA3.1-Myc-His-A-3)来源:invitrogen4)用途:真核核表达4))质粒图谱登记号:00252)质粒名称:pSUPER.neo3)来源:4)用途:siRNA5)是否可以提供更详细资料:可以6)是否可以共享:交换7)联系方式:PM)质粒图谱登记号:00312)质粒名称:pSilencer1.0-siRNA3)来源:Ambion4)用途:RNAi5)是否可以提供更详细资料:/techlib/Documents.html?fkResSxn=7&fkSub Sxn=236)是否可以共享:实验结束后,可提供含shRNA模板的质粒7)联系方式:pm5) )质粒图谱登记号:00322)质粒名称:pSilencer2.0-U6siRNA 3)来源:Ambion4)用途:RNAi ,与1.0相比,可以建立稳转株5)更详细资料:/techlib/prot/fm_7209.pdf 6)是否可以共享:交换 7)联系方式:pm6)会员名:mlluoE-mail:*************可提供试验资源名称和简要介绍:pSilencer 3.1-H1 neo Vector,是Ambion公司目前最高版本的shRNA 载体,为扩增此载体我已插入目的片段,如有战友需要,将此片段双酶切再连上自己的片段即可。
(转载)一. 九种表达载体Pllp-OmpA, pllp-STII, pMBP-P, pMBP-C,pET-GST, pET-Trx, pET-His, pET-CKS, pET-DsbA二. 克隆载体pTZ19RDNApUC57DNAPMD18TPQE30pUC18pUC19pTrcHisApTrxFuspRSET-ApRSET-BpVAX1PBR322pbv220pBluescriptIIKS( )L4440pCAMBIA-1301pMAL-p2XpGD926三.PET 系列表达载体ProteinExpression?ProkaryoticExpression?pETDsbFusionSystems39band40b ProteinExpression?ProkaryoticExpression?pETExpressionSystem33b ProteinExpression?ProkaryoticExpression?pETExpressionSystems ProteinExpression?ProkaryoticExpression?pETExpressionSystemsplusCompetentCells ProteinExpression?ProkaryoticExpression?pETGSTFusionSystems41and42 ProteinExpression?ProkaryoticExpression?pETNusAFusionSystems43.1and44 ProteinExpression?ProkaryoticExpression?pETVectorDNAProteinPurification?PurificationSystems?Strep?TactinResinsandPurificationKits四.PGEX 系列表达载体TEcoR?pGEX-1I/BAPpGEX-2TpGEX-2TKpGEX-3XpGEX-4T-1pGEX-4T-2pGEX-4T-3pGEX-5X-1pGEX-5X-2pGEX-5X-3pGEX-6P-1pGEX-6P-2pGEX-6P-3五.PTYBsystemPTYB1PTYB2PTYB11PTYB12六. 真核表达载体pCDNA3.1(-)pCDNA3.1( )pPICZalphaApGAPZα APYES2.0pBI121pEGFP-N1pEGFP-C1pPIC9KpPIC3.5K如何阅读分析质粒图谱载体主要有病毒和非病毒两大类, 其中质粒DNA是一种新的非病毒转基因载体。
基因酷质粒图谱/bbs/forum-38-1.html,收藏了将近800种质粒的图谱及相关信息特向大家推荐,介绍及使用方法见:/bbs/thread-417-1-1.html质粒图谱信息一.九种表达载体Pllp-OmpA, pllp-STII, pMBP-P, pMBP-C,pET-GST, pET-Trx, pET-His, pET-CKS, pET-DsbA二.克隆载体pTZ19R DNApUC57 DNAPMD18TPQE30pUC18pUC19pTrcHisApTrxFuspRSET-ApRSET-BpVAX1PBR322pbv220pBluescript II KS (+)L4440pCAMBIA-1301pMAL-p2XpGD926三.PET系列表达载体Protein Expression » Prokaryotic Expression » pET Dsb Fusion Systems 39b and 40bProtein Expression » Prokaryotic Expression » pET Expression System 33b Protein Expression » Prokaryotic Expression » pET Expression Systems Protein Expression » Prokaryotic Expression » pET Expression Systems plus Competent CellsProtein Expression » Prokaryotic Expression » pET GST Fusion Systems 41 and 42Protein Expression » Prokaryotic Expression » pET NusA Fusion Systems 43.1 and 44Protein Expression » Prokaryotic Expression » pET Vector DNAProtein Purification » Purification Systems » Strep•Tactin Resins and Purification Kits四.PGEX系列表达载体T EcoR pGEX-1 I/BAPpGEX-2TpGEX-2TKpGEX-3XpGEX-4T-1pGEX-4T-2pGEX-4T-3pGEX-5X-1pGEX-5X-2pGEX-5X-3pGEX-6P-1pGEX-6P-2pGEX-6P-3五.PTYB systemPTYB1PTYB2PTYB11PTYB12六.真核表达载体pCDNA3.1(-)pCDNA3.1(+)pPICZ alpha ApGAPZαAPYES2.0pBI121pEGFP-N1pEGFP-C1pPIC9KpPIC3.5K如何阅读分析质粒图谱载体主要有病毒和非病毒两大类,其中质粒DNA是一种新的非病毒转基因载体。
酿酒酵母表达载体pYES2,pYES2/NT,pYES2/CT,pYES3,pYES6, pYCplac22-GFP,酵母载体pAUR123,pRS303TEF,pRS304, pRS305,pRS306,pY13TEF,pY14TEF,pY15TEF,pY16TEF,酵母基因重组表达载体pUG6, pSH47,酵母单杂载体pHISi,pLacZi,pHIS2, pGAD424, 酵母双杂交系统:酿酒酵母Y187, 酿酒酵母AH109;质粒pGADT7,pGBKT7;对照质粒pGBKT7-53,pGBKT7-lam,pGADT7-T,PCL1,酿酒酵母菌株INVSc1,YM4271, AH109,Y187,Y190,毕赤酵母表达载体pPIC9K,pPIC9K-His,pPIC3.5K,pPICZalphaA,B,C,pPICZA,B,C,pGAPZαA,pAO815,pPIC9k-His,pHIL-S1,pPink hc,配套毕赤酵母Pichiapink,毕赤酵母宿主X33,KM71,KM71H,GS115,原核表达载体pQE30,31,32,40,60,61,62,等原核表达载体,包括pET系列,pET-GST,pGEX系列(含GST标签),pMAL系列pMAL-c2x,-c4x,-c4e,-c5x,-p5x,pBAD,pBADHis,pBADmycHis系列,pQE系列,pTrc99a,pTrcHis系列,pBV220,221,222,pTXB系列,pLLP-ompA,pIN-III-ompA (分泌型表达系列),pQBI63(原核表达带荧光)pET3a, pET 3d, pET 11a, pET 12a, pET 14b, pET 15b, pET 16b, pET 17b, pET 19b, pET 20b, pET 21a,b,d, pET 22b, pET 23a, pET 23b, pET 24a,b, pET 25b, pET 26b, pET 27b, pET 28a,b, pET 29a, pET 30a, pET 31b, pET 32a, pET 35b, pET 38b, pET 39b, pET 40b, pET 41a,b pET 42a, pET 43.1a,b pET 44a, pET 49b pET302,303 pET His,pET Dsb,pET GST,pET Trx pQE2, pQE9 pQE30,31,32, pQE 40 pQE70 pQE80L pQETirs system pRSET-A pRSET-B pRSET-C pGEX4T-1,-2,-3,5x-1,6p-1,6p-2,2tk,3c pBV220,221,222 pTrcHisA,B,C pBAD24,34,43 pBAD HisA,B,C pPinPoint-Xa1,Xa2,Xa3 pMALc2x, p2x pBV220 pGEM Ex1, pGEM7ZF(+), pTrc99A, pTwin1, pEZZ18 pkk232-8,pkk233-3,pACYC184,pBR322,pUC119 pTYB1,pTYB2,pTYB4,pTYB11 pBlueScript SK (+),pBlueScript SK(-)pLLP ompA, pINIIIompA, pMBP-P ,pMBP-C, 大肠杆菌冷激质粒: pColdI pColdII pColdIII pColdTF 原核共表达质粒:..pACYCduet-1,pETduet-1,pCDFduet-1,pRSFduet-1 Takara公司大肠杆菌分子伴侣: pG-KJE8 pGro7 pKJE7 pGTf2 pTf16 大肠杆菌宿主细胞: DH5a JM101 JM103 JM105 JM107 JM109 JM110 Top10 Top10F BL21(DE3)HB101 ER2529 E2566 C2566 MG1655 XL-10gold XL blue M15 JF1125 K802 SG1117 BL21(AI)BL21(DE3)plysS TG1 TB1 DH5a(pir)Tuner(DE3)Bl21 codonplusRIPL Novablue (DE3)Rosetta Rosetta(DE3)Rosetta(DE3)plys Rosetta-gami(DE3)Rosetta-gamiB(DE3), Rosetta-gamiB(DE3)plysS Orgami(DE3)OrgamiB(DE3)HMS174(DE3)植物表达/RNAi载体农杆菌pBI121,pBI121-GFP,pBI101,pBI221,pSN1301,pUN1301,pRTL2 , pRTL2-GFP , pRTL2-CFP, pRTL2-RFP , pRTL2-YFP,pCAMBIA 1300, 1301, 1302,1303,1304,1305, 1381Z,1391Z,2300, 2301,3300,3301,pCAMBIA super1300,pCAMBIAsuper1300-GFP,pPZP212,pPZP2121,pPZP212-GFP,pGDG,RNAi载体pART27,pHANNIBAL,pKANNIBAL, pFGC5941,pTCK303, pTRV1,pTRV2,T-DNA插入载体(随机突变体库)pSKI015,pSKI074,真菌ATMT载体pBIG2RHPH2-GUS-GFP,pBHt1枯草芽孢杆菌表达载体pWB980,pHT43,pHP13,pHP43,pBE2,pMUTIN4,pUB110,pE194,pMA5, pMK3,pMK4,pHT304,pHY300PLK,pBest502,pDG1363,pSG1154,pAX01, pSAS144,pDL,pDG148-stu,pDG641,pAL12,pUCX05-bgaB,pHT01,配套菌株BS 168,WB600,WB800,WB700,WB800N,1012,FZB42,1A747,广宿主质粒pVLT33RNAi基因沉默干扰敲除载体pSilencer1.0,pSilencer 2.1-U6 hygro, pSilencer 3.1-H1 hygro,pSilencer 3.1-H1 neo, pSilencer 4.1-CMV neo, pSilencer 4.1-CMV puro pMIR-REPORT Luciferase RNAi载体(oligoengine)pSuper-puro RNAi逆转录病毒载体(clontech): RNAi-Ready pSIREN-Retro Q, RNAi-ReadypSIREN-RetroQ-ZsGreen(Luciferase shRNA Annealed Oligonucleotide)RNAi慢病毒载体(addgene): pLKO.1..哺乳动物表达载体pcDNA3.1+/-,pcDNA4/HisMax B,pSecTag2 A,pVAX1,pBudCE4.1,pTracer CMV2,pcDNA3.1(-)/myc-His A ,pcDNA6-Myc/His B,pCEP4, pIRES,pIRESneo,pIRES hyg3,pCMV-myc,pCMV-HA,pIRES-puro3,pIRES-neo3,pCAGGS哺乳动物双杂交系统pACT,pBIND,pACT-MyoD,pBIND-Id,pG5luc,pCMV-BD, pCMV-AD, pBD-p53, pFR-luc,Cytotrap Two-Hybrid System:pSos, pSos MAFB, pMyr蜕皮激素诱导系统pIND, pVgRxR,LacSwith II哺乳动物诱导表达系统:pOPRSVI ,pOPI3CAT,pCMVLacI,GeneSwitch System:pSwitch哺乳动物表面展示系统:pDisplay, 四环素调控系统(Invitrogen):pcDNA4/TO/Myc-His A,pcDNA4/TO/Myc-His B,pcDNA4/TO/Myc-His C,pcDNA4/TO/Myc-His/LacZ,pcDNA6/TR四环素调控系统(Clontech):pTet-On,pTet-Off,pTRE2,pRevTRE,pRevTet-On,pRevTet-off信号通路报告载体:pGAS-TA-Luc,pSTAT3-TA-Luc, pISRE-TA-Luc, pTA-Luc,pIκB-EGFP,pNFAT-TA-Luc,pCaspase3-sensor,pAP1(PMA)-Luc;pGL4.26[luc2P/minP/Hygro],pGL4.29[luc2P/CRE/Hygro],pGL4.30[luc2P/NFA T-RE/Hygro],pGL4.75;p53-Luc,pAP-1-Luc, pNF-κB-Luc,pSRE-Luc,pFA2-Elk1,pFC-MEKK,pFR-luc,Gateway系统(invitrogen)pcDNA6.2-GWEmGFP-miR negative, pLenti 6/TR,pcDNA 6.2-GW EmGFP-miR,乳酸菌表达载体及各种乳酸菌乳酸杆菌菌株,pNZ8148,pLEISS,pMG36e,pBBR1MCS-5,pBBR1MCS-6,pRV610,pLEM415,pHY3 00PLK,分泌型乳酸菌表达载体pVE5523,pPG611.1,pPG612.1等和乳酸杆菌菌株宿主菌NZ9000,MG1363,Lactobacillus casei 1.539,Lactobacillus casei,acidophilus NCFM,1.2,Lactobacillus sakei 23K,L.plantarum,L.rhamnosusGG,B.coagulans,Bifidobacterium bifidum,Bifidobacterium infantis,Lactococcus lactis M17,1663,Lactobacillus reuterii广宿主表达载体链球菌表达敲除载体假单胞菌表达载体pVLT33,pBBR1MCS-2,3,4,5,6, pJRD215,pJN105,pME6032,Cos载体pLAFR3,pMP2444(GFP), pHY300PLK,pRT102,pRL1063a, 转座子载体pUT-miniTn5,pMGS100, pWHM10,pKC1139,pSET152,pOJ260,pPG611.1,pPG612.1,..腺病毒载体/慢病毒,逆转录病毒表达载体及包装包膜质粒,腺病毒系统(Stratagene): pAdEasy-1,pShuttle-CMV,pShuttle,pAdTrack, pAdTrack-CMV, pShuttle-IRES-hrGFP-1、pShuttle-IRES-hrGFP-2、pShuttle-CMV-lacZ,pShuttle-CMV-EGFP-C,pXC1, pBHGE3, 配套大肠杆菌BJ5183,293,293T cellline 腺相关病毒系统(Stratagene): pAAV-MCS,pAAV-RC,pHelper,pAAV-LacZ,pAAV-IRES-hrGFP,pCMV-MCS,慢病毒载体:pLVX-DsRed-Monomer-N1,pLVX-IRES-ZsGreen1,pLVX-AcGFP1-N1,Lenti6/v 5-EDST-EGFP,pWPXL, FUGW,pLentilox 3.7,RNAi-Ready pSIREN-Retro Q,RNAi-Ready pSIREN-Retro Q-ZsGreen,pSUPER.Retro-GFP/Neo,pSUPER-Retro-Neo, pSUPER.Retro-puro,PLNCX PLNCX2 pMSCV-HYG pMSCV-neo pMSCV-puro pLEGFP-C1 pLOX-CW-CRE pLOX-GFP-IRES-TK pRetroX-IRES-DsRedExpress, pLVX-IRES-mCherry质粒载体。
实验新⼈必读(七):⼀⽂学会读懂和查找质粒图谱作者:解螺旋.⼦⾮鱼如需转载请注明来源:解螺旋·医⽣科研助⼿导语质粒图谱即为质粒DNA序列的物理图谱,包含了质粒⼤⼩、筛选标记、克隆位点、转录及翻译元件等信息。
尽管它为我们选择质粒、了解质粒特点及应⽤提供了重要依据,然⽽我们常常要为图谱中丰富信息所困扰。
那么,本⽂就为你拨云见⽇,让你快速掌握质粒图谱。
三步法看懂质粒图谱Step 1:先了解质粒的基本组成元素。
1)复制起始点Ori。
该位点决定了质粒的宿主及质粒的拷贝数,它是质粒中⼀段特定序列,富含AT和重复序列。
Tips:图谱上只有⼀个Ori,表⽰质粒是原核克隆表达质粒;有两个Ori,则表⽰该质粒是,穿梭质粒,即可在原核也可在真核中复制。
2)抗性筛选基因。
图谱中Kan/tet就是抗⽣素抗性基因,⽅便后续通过抗⽣素筛选阳性克隆。
特点就是单词最后会以⼤写R或上标r结束。
Tips:⼀般克隆载体只有⼀种抗性筛选标记,部分表达载体及穿梭质粒具有两种抗性筛选标记。
3)多克隆位点(MCS),即⼀系列限制性内切酶酶切位点,是外源DNA插⼊位点,⼀般可通过酶切/连接⽅式将外源DNA插⼊质粒,外源DNA⼀般⼩于10kb,⽽⽚段越长,转化效率越低。
Tips: ⼀般位于转录启动和转录终⽌信号之间;所包含的限制性内切酶位点数量和组成因载体不同会有所差异,且其中的酶切位点在质粒中为单⼀的酶切位点;同时在使⽤时需注意质粒载体与外源DNA酶切位点的兼容性问题4)荧光标记或蛋⽩标签序列。
蛋⽩纯化标签蛋⽩:His-Tag,GST-Tag等;蛋⽩检测标签蛋⽩:Myc-Tag,Flag-Tag,HA-Tag等;荧光蛋⽩表达标签:GFP,mCherry等。
Step 2:看质粒是否是表达载体,如果是,那就必定有这些原件:启动⼦-核糖体结合位点-克隆位点-转录终⽌信号。
1、启动⼦:促进DNA转录的DNA序列,可与RNApol特异性结合。
2、增强⼦/沉默⼦:前者是真核基因组中⼀种增强邻近基因转录过程的调控顺序,其作⽤与增强⼦所在位置或⽅向⽆关。
Restriction Map and Multiple Cloning Site (MCS) of pECFP-N1. All restriction sites shown are unique.pECFP-N1 Vector Information PT3285-5Catalog #6900-1(PR29991; published 04 October 2002)DescriptionpECFP-N1 encodes an enhanced cyan fluorescent variant of the Aequorea victoria green fluores-cent protein gene (GFP). The ECFP gene contains six amino acid substitutions. The Tyr-66 to Trp substitution gives ECFP fluorescence excitation (major peak at 433 nm and a minor peak at 453 nm)and emission (major peak at 475 nm and a minor peak at 501 nm) similar to other cyan emission variants (1, 2). The other five substitutions (Phe-64 to Leu; Ser-65 to Thr; Asn-146 to Ile; Met-153to Thr; and Val-163 to Ala) enhance the brightness and solubility of the protein, primarily due to improved protein folding properties and efficiency of chromophore formation (1, 3, 4). In addition to the chromophore mutations, ECFP contains >190 silent mutations that create an open reading frame comprised almost entirely of preferred human codons (5, 6). Furthermore, upstream sequences flanking ECFP have been converted to a Kozak consensus translation initiation site (7). These changes increase the translational efficiency of the ECFP mRNA and consequently the expression of ECFP in mammalian and plant cells.The vector contains an SV40 origin of replication and a neomycin resistance (Neo r ) gene for selection (using G418) in mammalian cells. A bacterial promoter upstream of this cassette (P ) expresses kanamycin resistance in E. coli . The vector backbone also provides a pUC19 origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production. The MCS in pECFP-N1 is between the immediate early promoter of CMV (P CMV IE ) and the ECFP coding sequences. Genes cloned into the MCS will be expressed as fusions to the N-terminus of ECFP if they are in the same reading frame as ECFP and there are no intervening stop codons. The inserted gene should include an initiating ATG codon. ECFP fusion proteins retain the fluorescent properties of the native protein in vivo .UseThe recombinant ECFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (8). pECFP-N1 can also be used simply to express ECFP in a cell line of interest (e.g., as a transfection marker). ECFP can be used for double-labeling experiments together with EYFP using standard fluorescence microscopy and the appropriate filter sets.G CTA GCG CTA CCG GAC TCA GAT CTC GAG CTC AAG CTT CGA ATT CTG CAG TCG ACG GTA CCG CGG GCC CGG GAT CCA CCG GTC GCC ACC 591•611•601•621•631•641•661•651•671•Hin d III Xho I Apa I Bsp 120 I Kpn I Asp 718 I Bam H I ECFPAge I Xma I Sma ISac II Sal I Acc I Sac I Ecl 136 II Eco R I Pst I Eco 47 IIIBgl II Nhe II (1402) (1640)Eco (3856)(2579)Ase IG I (1389)Location of features•Human cytomegalovirus (CMV) immediate early promoter: 1–589Enhancer region: 59–465TATA box: 554–560Transcription start point: 583C→G mutation to remove Sac I site: 569•MCS: 591–671•Enhanced cyan fluorescent protein (ECFP) geneKozak consensus translation initiation site: 672–682Start codon (ATG): 679–681; stop codon: 1396–1398Insertion of Val at position 2: 682–684ECFP mutations:Phe-64 to Leu, Ser-65 to Thr, and Tyr-66 to Trp: 871–879Asn-146 to Ile: 1117–1119Met-153 to Thr: 1138–1140Val-163 to Ala: 1168–1170His-231 to Leu mutation (A→T): 1373•SV40 early mRNA polyadenylation signalPolyadenylation signals: 1552–1557 & 1581–1586mRNA 3' ends: 1590 & 1602•f1 single-strand DNA origin: 1649–2104(Packages the noncoding strand of ECFP.)•Bacterial promoter for expression of Kan r gene:–35 region: 2166–2171; –10 region: 2189–2194Transcription start point: 2201•SV40 origin of replication: 2445–2580•SV40 early promoterEnhancer (72-bp tandem repeats): 2278–2349 & 2350–242121-bp repeats: 2425–2445, 2446–2466 & 2468–2488Early promoter element: 2501–2507Major transcription start points: 2497, 2535, 2541 & 2546•Kanamycin/neomycin resistance geneNeomycin phosphotransferase coding sequences:Start codon (ATG): 2629–2631; stop codon: 3421–3423G→A mutation to remove Pst I site: 2811C→A (Arg to Ser) mutation to remove Bss H II site: 3157•Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signalPolyadenylation signals: 3659–3664 & 3672–3677•pUC plasmid replication origin: 4008–4651Primer Locations•EGFP-N Sequencing Primer (#6479-1): 745–724•EGFP-C Sequencing Primer (#6478-1): 1332–1353Propagation in E. coli•Suitable host strains: DH5α, HB101, and other general-purpose strains. Single-stranded DNA production requires a host containing an F plasmid such as JM101 or XL1-Blue.•Selectable marker: plasmid confers resistance to kanamycin (30 µg/ml) in E. coli hosts.• E. coli replication origin: pUC;Copy number: ~500•Plasmid incompatibility group: pMB1/Col E1References1.Heim, R.,& Tsien, R. Y. (1996) Curr. Biol. 6:178–182.2.Heim, R., et al. (1994) Proc. Natl. Acad. Sci. USA 91:12501–12504.3.Cormack, B., et al. (1996) Gene 173:33–38.4.Heim, R., et al. (1995) Nature373:663–664.5.Yang, T. T., et al. (1996) Nucleic Acids Res.24:4592–4593.6.Haas, J., et al. (1996) Curr. Biol.6:315–324.7.Kozak, M. (1987) Nucleic Acids Res.15:8125–8148.8.Gorman, C. (1985) In DNA cloning: A practical approach, vol. II. Ed. D. M. Glover. (IRL Press, Oxford, U.K.), pp. 143–190.BD Biosciences Clontech Protocol # PT3285-5 2Version # PR29991Protocol # PT3285-5 BD Biosciences ClontechVersion # PR299913Notice to PurchaserUse of BD Biosciences Clontech’s Living Colors™ products containing DNA sequences coding for mutant Aequorea victoria green fluorescent protein (GFP) variants or proteins thereof requires a license from Amersham Biosciences under U.S. Patent Nos. 5,625,048; 5,777,079; 6,054,321and other pending U.S. and foreign patent applications. In addition, certain BD Biosciences Clontech products are made under U.S. Patent No.5,804,387 licensed from Stanford University.Not-For-Profit research institutes or entities are granted an automatic license with the purchase of this product for use in non-commercial internal research purposes, the terms of which are disclosed in detail in the license that accompanies the shipment of this product. Such license specifi-cally excludes the right to sell or otherwise transfer this product or its components to third parties.For-Profit research institutes or entities must obtain a license from Amersham Biosciences. E-mail: gfp@Please contact BD Biosciences Clontech directly for any other assistance, including purchasing and technical support. All companies and institutions purchasing Living Colors™ products will be included in a quarterly report to Aurora Biosciences, as required by the BD Biosciences Clontech/Aurora Biosciences license agreement.This product is intended to be used for research purposes only. It is not to be used for drug or diagnostic purposes nor is it intended for human use. BD Biosciences Clontech products may not be resold, modified for resale, or used to manufacture commercial products without written approval of BD Biosciences Clontech.© 2002, Becton, Dickinson and Company Note: The attached sequence file has been compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by BD Biosciences Clontech. This vector has not been completely sequenced.。