pLVX-Puro慢病毒载体使用说明_精品文档.pdf

pLVX-Puro慢病毒载体使用说明_精品文档.pdf

《pLVX-Puro慢病毒载体使用说明_精品文档.pdf》由会员分享，可在线阅读，更多相关《pLVX-Puro慢病毒载体使用说明_精品文档.pdf（7页珍藏版）》请在冰豆网上搜索。

pLVX-PuropLVX-Puro载体基本信息载体基本信息：

载体名称载体名称:

pLVX-Puro,pLVXpuro质粒类型质粒类型:

哺乳动物细胞慢病毒表达载体哺乳动物细胞慢病毒表达载体高拷贝高拷贝/低拷贝低拷贝:

高拷贝高拷贝启动子启动子:

CMV克隆方法克隆方法:

多克隆位点，限制性内切酶多克隆位点，限制性内切酶载体大小载体大小:

8102bp5测序引物及序列测序引物及序列:

CMV-F：

CGCAAATGGGCGGTAGGCGTG3测序引物及序列测序引物及序列:

-载体标签载体标签:

-载体抗性载体抗性:

氨苄氨苄筛选标记筛选标记:

嘌呤霉素嘌呤霉素备注备注:

含有组成型含有组成型CMV启动子的慢病毒载体启动子的慢病毒载体稳定性稳定性:

/组成型组成型:

-病毒病毒/非病毒非病毒:

慢病毒慢病毒pLVX-Puro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：

编号编号载体名称载体名称北京华越洋北京华越洋VECT231322pLVX-PuropLVX-Puro载体载体简介：

简介：

DescriptionpLVX-PuroisanHIV-1-based,lentiviralexpressionvector.Lentiviralparticlesderivedfromthevectorallowyoutoexpressyourgeneofinterestinvirtuallyanycelltype,evenprimarycells.Expressionofyourgeneisdrivenbytheconstitutivelyactivehumancytomegalovirusimmediateearlypromoter（PCMVIE）,locatedjustupstreamofthemultiplecloningsite（MCS）,allowingconstitutive,highlevelexpressionofyourproteinofinterest.pLVX-Purocontainsalloftheviralprocessingelementsnecessaryfortheproductionofreplication-incompetentlentivirus,aswellaselementstoimproveviraltiter,transgeneexpression,andoverallvectorfunction.Thewoodchuckhepatitisvirusposttranscriptionalregulatoryelement（WPRE）promotesRNAprocessingeventsandenhancesnuclearexportofviralandtransgeneRNA

（1）,leadingtoincreasedviraltitersfrompackagingcells,andenhancedexpressionofyourgeneofinterestintargetcells.Inaddition,thevectorincludesaRev-responseelement（RRE）,whichfurtherincreasesviraltitersbyenhancingthetransportofunsplicedviralRNAoutofthenucleus

（2）.Finally,pLVX-Puroalsocontainsacentralpolypurinetract（cPPT）elementthatincreasesnuclearimportationoftheviralgenomeduringtargetcellinfection,resultinginimprovedvectorintegrationandmoreefficienttransduction（3）.Inadditiontolentiviralelements,pLVX-Purocontainsapuromycinresistancegene（Puror）underthecontrolofthemurinephosphoglyceratekinase（PGK）promoter（PPGK）fortheselectionofstabletransductants.ThevectoralsocontainsapUCoriginofreplicationandanE.coliampicillinresistancegene（Ampr）forpropagationandselectioninbacteria.UsepLVX-PuroconstitutivelyexpressesyourgeneofinterestfromPCMVIEwhentransducedintotargetcells.Beforethevectorcanbetransducedintocells,however,itmustbetransfectedinto293TpackagingcellswithourLenti-XHTPackagingSystem（Cat.Nos.632160and632161）.Thispackagingsystemallowsyoutosafelyproducehightiter,infectious,replication-incompetent,VSV-Gpseudotypedlentiviralparticlesthatcaninfectawiderangeofcelltypes,includingnon-dividingandprimarycells（4）.pLVX-Puro载体序列载体序列：

ORIGIN1TGGAAGGGCTAATTCACTCCCAAAGAAGACAAGATATCCTTGATCTGTGGATCTACCACA61CACAAGGCTACTTCCCTGATTAGCAGAACTACACACCAGGGCCAGGGGTCAGATATCCAC121TGACCTTTGGATGGTGCTACAAGCTAGTACCAGTTGAGCCAGATAAGGTAGAAGAGGCCA181ATAAAGGAGAGAACACCAGCTTGTTACACCCTGTGAGCCTGCATGGGATGGATGACCCGG241AGAGAGAAGTGTTAGAGTGGAGGTTTGACAGCCGCCTAGCATTTCATCACGTGGCCCGAG301AGCTGCATCCGGAGTACTTCAAGAACTGCTGATATCGAGCTTGCTACAAGGGACTTTCCG361CTGGGGACTTTCCAGGGAGGCGTGGCCTGGGCGGGACTGGGGAGTGGCGAGCCCTCAGAT421CCTGCATATAAGCAGCTGCTTTTTGCCTGTACTGGGTCTCTCTGGTTAGACCAGATCTGA481GCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCT541TGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTC601AGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAG661CGAAAGGGAAACCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGG721CAAGAGGCGAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGA781AGGAGAGAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGG841AAAAAATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGG901CAAGCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCT961GTAGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGAT1021CATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACA1081CCAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAGC1141AAGCGGCCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAG1201TGAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGC1261AAAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTGG1321GTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGC1381CAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGC1441GCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCT1501GGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCTCTGGAAA1561ACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACA1621GATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTT1681AATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATT1741GGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGGCTGTGGTA1801TATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGT1861ACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCT1921CCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGA