pET-SUMO大肠杆菌表达载体说明.pdf

pET-SUMO大肠杆菌表达载体说明.pdf

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pET-SUMO编号编号载体名称载体名称北京华越洋生物VECT4790pET-SUMOpETsumo载体基本信息载体基本信息载体名称载体名称:

pET-SUMO质粒类型质粒类型:

大肠杆菌表达载体高拷贝高拷贝/低拷贝低拷贝:

低拷贝启动子启动子:

T7和lacO克隆方法克隆方法:

TACloning载体大小载体大小:

5643bp5测序引物及序列测序引物及序列:

T7Forward3测序引物及序列测序引物及序列:

-载体标签载体标签:

HisTag（6x）；SUMOTag载体抗性载体抗性:

卡那霉素筛选标记筛选标记:

-备注备注:

宿主菌株BL21（DE3）；IPTG诱导稳定性稳定性:

-组成型组成型:

诱导型病毒病毒/非病毒非病毒:

非病毒pETsumo载体质粒图谱和多克隆位点信息载体质粒图谱和多克隆位点信息pETsumo载体简介载体简介TheChampionpET-SUMOExpressionSystemproducesthehighestlevelsofsolubleproteininE.coli.Itutilizesasmallubiquitin-relatedmodifier（SUMO）fusion,belongingtothegrowingfamilyofubiquitin-relatedproteins,toenhancethesolubilityofexpressedfusionproteins.Incontrasttoubiquitin,SUMOisinvolvedinthestabilizationandlocalizationofproteinsinvivo.Afterexpression,the11kdSUMOmoietycanbecleavedbythehighlyspecificandactiveSUMO（ULP-1）proteaseatthecarboxylterminal,producinganativeprotein*.TheChampionpETSUMOProteinandPeptideExpressionSystemoffers:

GreatlyenhancedsolubilitywithanN-terminalSUMOfusionHighlyefficientcleavage-producesnativeproteinofinterestwithSUMO（ULP-1）protease*Highlyspecificcleavage-eliminatesthechanceofyourproteinofinterestbeinginternallydigested,regardlessofitsaminoacidsequenceSignificantlyincreasedstabilitywithSUMOfusion-canbeusedforsmallpeptideproductionT7lacpromoterforhigh-levelproteinexpressionN-terminal6xHistagforproteindetectionandpurification.pETsumo载体序列载体序列CAAGGAGATGGCGCCCAACAGTCCCCCGGCCACGGGGCCTGCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCGAAGTGGCGAGCCCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCAGCAACCGCACCTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTAGAGGATCGAGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGGGCAGCAGCCATCATCATCATCATCACGGCAGCGGCCTGGTGCCGCGCGGCAGCGCTAGCATGTCGGACTCAGAAGTCAATCAAGAAGCTAAGCCAGAGGTCAAGCCAGAAGTCAAGCCTGAGACTCACATCAATTTAAAGGTGTCCGATGGATCTTCAGAGATCTTCTTCAAGATCAAAAAGACCACTCCTTTAAGAAGGCTGATGGAAGCGTTCGCTAAAAGACAGGGTAAGGAAATGGACTCCTTAAGATTCTTGTACGACGGTATTAGAATTCAAGCTGATCAGACCCCTGAAGATTTGGACATGGAGGATAACGATATTATTGAGGCTCACAGAGAACAGATTGGTGGTAGACAAGCTTAGGTATTTATTCGGCGCAAAGTGCGTCGGGTGATGCTGCCAACTTAGTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATTGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAATTAATTCTTAGAAAAACTCATCGAGCATCAAATGAAACTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTTCCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAATAAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCAGACTTGTTCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGACGAAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAATGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAATATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAATGCTGTTTTCCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGGAGTACGGATAAAATGCTTGATGGTCGGAAGAGGCATAAATTCCGTCAGCCAGTTTAGTCTGACCATCTCATCTGTAACATCATTGGCAACGCTACCTTTGCCATGTTTCAGAAACAACTCTGGCGCATCGGGCTTCCCATACAATCGATAGATTGTCGCACCTGATTGCCCGACATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTAGAGCAAGACGTTTCCCGTTGAATATGGCTCATAACACCCCTTGTATTACTGTTTATGTAAGCAGACAGTTTTATTGTTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCAATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGGCAGCTGCGGTAAAGCTCATCAGCGTGGTCGTGAAGCGATTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTCTCCAGAAGCGTTAATGTCTGGCTTCTGATAAAGCGGGCCATGTTAAGGGCGGTTTTTTCCTGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGTAATGATACCGATGAAACGAGAGAGGATGCTCACGATACGGGTTACTGATGATGAACATGCCCGGTTACTGGAACGTTGTGAGGGTAAACAACTGGCGGTATGGATGCGGCGGGACCAGAGAAAAATCACTCAGGGTCAATGCCAGCGCTTCGTTAATACAGATGTAGGTGTTCCACAGGGTAGCCAGCAGCATCCTGCGATGCAGATCCGGAACATAATGGTGCAGGGCGCTGACTTCCGCGTTTCCAGACTTTACGAAACACGGAAACCGAAGACCATTCATGTTGTTGCTCAGGTCGCAGACGTTTTGCAGCAGCAGTCGCTTCACGTTCGCTCGCGTATCGGTGATTCATTCTGCTAACCAGTAAGGCAACCCCGCCAGCCTAGCCGGGTCCTCAACGACAGGAGCACGATCATGCGCACCCGTGGGGCCGCCATGCCGGCGATAATGGCCTGCTTCTCGCCGAAACGTTTGGTGGCGGGACCAGTGACGAAGGCTTGAGCGAGGGCGTGCAAGATTCCGAATACCGCAAGCGACAGGCCGATCATCGTCGCGCTCCAGCGAAAGCGGTCCTCGCCGAAAATGACCCAGAGCGCTGCCGGCACCTGTCCTACGAGTTGCATGATAAAGAAGACAGTCATAAGTGCGGCGACGATAGTCATGCCCCGCGCCCACCGGAAGGAGCTGACTGGGTTGAAGGCTCTCAAGGGCATCGGTCGAGATCCCGGTGCCTAATGAGTGAGCTAACTTACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCCAGGGTGGTTTTTCTTTTCACCAGTGAGACGGGCAACAGCTGATTGCCCTTCACCGCCTGGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCAGGCGAAAATCCTGTTTGATGGTGGTTAACGGCGGGATATAACATGAGCTGTCTTCGGTATCGTCGTATCCCACTACCGAGATATCCGCACCAACGCGCAGCCCGGACTCGGTAATGGCGCGCATTGCGCCCAGCGCCATCTGATCGTTGGCAACCAGCATCGCAGTGGGAACGATGCCCTCATTCAGCATTTGCATGGTTTGTTGAAAACCGGACATGGCACTCCAGTCGCCTTCCCGTTCCGCTATCGGCTGAATTTGATTGCGAGTGAGATATTTATGCCAGCCAGCCAGACGCAGACGCGCCGAGACAGAACTTAATGGGCCCGCTAACAGCGCGATTTGCTGGTGACCCAATGCGACCAGATGCTCCACGCCCAGTCGCGTACCGTCTTCATGGGAGAAAATAATACTGTTGATGGGTGTCTGGTCAGAGACATCAAGAAATAACGCCGGAACATTAGTGCAGGCAGCTTCCACAGCAATGGCATCCTGGTCATCCAGCGGATAGTTAATGATCAGCCCACTGACGCGTTGCGCGAGAAGATTGTGCACCGCCGCTTTACAGGCTTCGACGCCGCTTCGTTCTACCATCGACACCACCACGCTGGCACCCAGTTGATCGGCGCGAGATTTAATCGCCGCGACAATTTGCGACGGCGCGTGCAGGGCCAGACTGGAGGTGGCAACGCCAATCAGCAACGACTGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTGGGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCCCGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCACCACGCGGGAAACGGTCTGATAAGAGACACCGGCATACTCTGCGACATCGTATAACGTTACTGGTTTCACATTCACCACCCTGAATTGACTCTCTTCCGGGCGCTATCATGCCATACCGCGAAAGGTTTTGCGCCATTCGATGGTGTCCGGGATCTCGACGCTCTCCCTTATGCGACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTGAGGCCGTTGAGCACCGCCGCCGCAAGGAATGGTGCATG其他其他大肠杆菌大肠杆菌表达载体：

表达载体：

pBV221ptdTomatopET-52b（+）pAmCyanpDsRed-Express2pBV220pCold-GSTpColdS-SUMOpColdTFpColdIVpColdIIIpColdIIpColdIpE-SUMOpCold-ProS2pBAD102/D-TOPOpBAD202/D-TOPOpACYC184pBAD/Thio-TOPOpBad/Myc-HisCpBad/Myc-HisBpBad/Myc-HisApBad/HisCpBad/HisBpBad/HisApBAD-TOPOpET-23b（+）pET-23a（+）pET-23c（+）pET-23（+）pET-12b（+）pET-12c（+）pET-12a（+）pET-11b（+）pET-11a（+）pET-11c（+）pBad24pQE-82LpQE-81LpQE-80LpQE-32pQE-9pQE-16pQE-31pQE-60pQE-70pQE-40pET-51b（+）pET-50b（+）pET-49b（+）pET-48b（+）pET-47b（+）pET-26b（+）pET-32a（+）pET-21b（+）pET-22b（+）pET-14bpET-16bpET-15bpET-19bpET-20b（+）pET-21d（+）pET-21c（+）pET-21b（+）pET-21a（+）pET-24a（+）pET-24d（+）pET-25b（+）pET-27b（+）pET-28a（+）pET-30a（+）pET-42a（+）pET-43.1c（+）pET-43.1b（+）pET-43.1a（+）pET-44a（+）pET-44c（+）pET-46EK/LICpET-37b（+）pTrcHis2CpTrcHis2BpTrcHis2ApET303/CT-HispET302/NT-HispRSET-CFPpRSET-EmGFPpRSET-BFPpGFPuvpET300/NT-DESTpET301/CT-DESTpGEM-TpBad43pGEX-4T-3pGEX-5X-2pBlueScriptSK（+）pG-Tf2pG-KJE8pGro7pET-SUMOpSE380pET-17bpET102/D-TOPOpCDFDuet-1pMAL-p5xpTf16pET-28c（+）pBluescriptIISK（+）pET-30b（+）pSUMOpProEXHTcpProEXHTbpProEXHTapKD3pKD13pKD46pTYB1pTYB2pTWIN2pBluescriptIIKS（-）pTYB12pMAL-p5epACYCDuet-1pEGM-11ZF（+）pEGM-7ZF（+）PinPointXa-3PinPointXa-2PinPointXa-1pSP73pSP64pTWIN1pTYB11pTXB1pET-5b（+）pBad/gIIICpBad/gIIIBpBad/gIIIApET-5a（+）pMal-p4XpMal-p2Gpkk223-3pkk232-8pCYB1pEZZ18pBAD18pMAL-c5xpMal-p2EpMal-p2XpET-44EK/LICpET-43.1EK/LICpET-41EK/LICpMal-c4XpTrcHisBpET-31b（+）pET-3b（+）pET-41a（+）pGEX-3XpGEX-4T-2pETDuet-1pGEX-4T-1pTrc99apET-28b（+）pET-HispALEXa,b,cpACYC177pBR322pKD4pKD20pMXB10pEcoli-6xHN-GFPuvpKJE7pRSETBpGEX-KGpGEX-2TpRSFDuet-1pCOLADuet-1pTrcHisCpTrcHisApET-41b（+）pET-42b（+）pET-3a（+）pGEX-6P-3pGEX-6P-2pGEX-6P-1pGEX-5X-3pGEX-5X-1pGEX-2TKpRSETApMal-c2GpMal-c2EpMal-c2XpRSETCpQE-30pET-45b（+）pET-44b（+）pET-42c（+）pET-41c（+）pET-40b（+）pET-33b（+）pET-39b（+）pET-32EK/LICpET-32Xa/LICpET-32c（+）pET-32b（+）pET-30Xa/LICpET-30EK/LICpET-30c（+）pET-29c（+）pET-29b（+）pET-29a（+）pET-24c（+）pET-24b（+）pET-24（+）pET-23d（+）pET-11d（+）pBad33