pLVX-AcGFP1-N1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to AcGFP1, a green fl uorescent protein derived from Aequorea coerulescens. Genes cloned into the multiple cloning site (MCS), located upstream of the AcGFP1 coding sequence, are expressed as N-terminal fusions of the AcGFP1 protein. Expression of the fusion protein is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. Lentiviral particles derived from the vector allow the expression of AcGFP1 fusion proteins in virtually any cell type, including primary cells. The unmodifi ed vector expresses AcGFP1, and may be used to produce marker virus to optimize infection protocols.

pLVX-AcGFP1-N1 contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral and transgene RNA (1), leading to increased viral titers from packaging cells, and enhanced expression of your gene of interest in target cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (2). Finally, pLVX-AcGFP1-N1 also contains a central polypurine tract (cPPT) element that increases nuclear importation of the viral genome during target cell infection, resulting in improved vector integration and more effi cient transduction (3).

In addition to lentiviral elements, pLVX-AcGFP1-N1 contains a puromycin resistance gene (Puror ) under the control of the murine phosphoglycerate kinase (PGK) promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr ) for propagation and selection in bacteria.

載體應(yīng)用

pLVX-AcGFP1-N1 constitutively expresses your gene of interest from PCMV IE when transduced into target cells. Before the vector can be transduced into cells, however, it must be transfected into 293T packaging cells with our Lenti-X? HT Packaging System (Cat. Nos. 632160 and 632161). This packaging system allows you to safely produce high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including non-dividing and primary cells (4).

0001 TGGAAGGGCT AATTCACTCC CAAAGAAGAC AAGATATCCT TGATCTGTGG ATCTACCACA 
0061 CACAAGGCTA CTTCCCTGAT TAGCAGAACT ACACACCAGG GCCAGGGGTC AGATATCCAC 
0121 TGACCTTTGG ATGGTGCTAC AAGCTAGTAC CAGTTGAGCC AGATAAGGTA GAAGAGGCCA 
0181 ATAAAGGAGA GAACACCAGC TTGTTACACC CTGTGAGCCT GCATGGGATG GATGACCCGG 
0241 AGAGAGAAGT GTTAGAGTGG AGGTTTGACA GCCGCCTAGC ATTTCATCAC GTGGCCCGAG 
0301 AGCTGCATCC GGAGTACTTC AAGAACTGCT GATATCGAGC TTGCTACAAG GGACTTTCCG 
0361 CTGGGGACTT TCCAGGGAGG CGTGGCCTGG GCGGGACTGG GGAGTGGCGA GCCCTCAGAT 
0421 CCTGCATATA AGCAGCTGCT TTTTGCCTGT ACTGGGTCTC TCTGGTTAGA CCAGATCTGA 
0481 GCCTGGGAGC TCTCTGGCTA ACTAGGGAAC CCACTGCTTA AGCCTCAATA AAGCTTGCCT 
0541 TGAGTGCTTC AAGTAGTGTG TGCCCGTCTG TTGTGTGACT CTGGTAACTA GAGATCCCTC 
0601 AGACCCTTTT AGTCAGTGTG GAAAATCTCT AGCAGTGGCG CCCGAACAGG GACTTGAAAG 
0661 CGAAAGGGAA ACCAGAGGAG CTCTCTCGAC GCAGGACTCG GCTTGCTGAA GCGCGCACGG 
0721 CAAGAGGCGA GGGGCGGCGA CTGGTGAGTA CGCCAAAAAT TTTGACTAGC GGAGGCTAGA 
0781 AGGAGAGAGA TGGGTGCGAG AGCGTCAGTA TTAAGCGGGG GAGAATTAGA TCGCGATGGG 
0841 AAAAAATTCG GTTAAGGCCA GGGGGAAAGA AAAAATATAA ATTAAAACAT ATAGTATGGG 
0901 CAAGCAGGGA GCTAGAACGA TTCGCAGTTA ATCCTGGCCT GTTAGAAACA TCAGAAGGCT 
0961 GTAGACAAAT ACTGGGACAG CTACAACCAT CCCTTCAGAC AGGATCAGAA GAACTTAGAT 
1021 CATTATATAA TACAGTAGCA ACCCTCTATT GTGTGCATCA AAGGATAGAG ATAAAAGACA 
1081 CCAAGGAAGC TTTAGACAAG ATAGAGGAAG AGCAAAACAA AAGTAAGACC ACCGCACAGC 
1141 AAGCGGCCGG CCGCTGATCT TCAGACCTGG AGGAGGAGAT ATGAGGGACA ATTGGAGAAG 
1201 TGAATTATAT AAATATAAAG TAGTAAAAAT TGAACCATTA GGAGTAGCAC CCACCAAGGC 
1261 AAAGAGAAGA GTGGTGCAGA GAGAAAAAAG AGCAGTGGGA ATAGGAGCTT TGTTCCTTGG 
1321 GTTCTTGGGA GCAGCAGGAA GCACTATGGG CGCAGCGTCA ATGACGCTGA CGGTACAGGC 
1381 CAGACAATTA TTGTCTGGTA TAGTGCAGCA GCAGAACAAT TTGCTGAGGG CTATTGAGGC 
1441 GCAACAGCAT CTGTTGCAAC TCACAGTCTG GGGCATCAAG CAGCTCCAGG CAAGAATCCT 
1501 GGCTGTGGAA AGATACCTAA AGGATCAACA GCTCCTGGGG ATTTGGGGTT GCTCTGGAAA 
1561 ACTCATTTGC ACCACTGCTG TGCCTTGGAA TGCTAGTTGG AGTAATAAAT CTCTGGAACA 
1621 GATTTGGAAT CACACGACCT GGATGGAGTG GGACAGAGAA ATTAACAATT ACACAAGCTT 
1681 AATACACTCC TTAATTGAAG AATCGCAAAA CCAGCAAGAA AAGAATGAAC AAGAATTATT 
1741 GGAATTAGAT AAATGGGCAA GTTTGTGGAA TTGGTTTAAC ATAACAAATT GGCTGTGGTA 
1801 TATAAAATTA TTCATAATGA TAGTAGGAGG CTTGGTAGGT TTAAGAATAG TTTTTGCTGT 
1861 ACTTTCTATA GTGAATAGAG TTAGGCAGGG ATATTCACCA TTATCGTTTC AGACCCACCT 
1921 CCCAACCCCG AGGGGACCCG ACAGGCCCGA AGGAATAGAA GAAGAAGGTG GAGAGAGAGA 
1981 CAGAGACAGA TCCATTCGAT TAGTGAACGG ATCTCGACGG TATCGCCTTT AAAAGAAAAG 
2041 GGGGGATTGG GGGGTACAGT GCAGGGGAAA GAATAGTAGA CATAATAGCA ACAGACATAC 
2101 AAACTAAAGA ATTACAAAAA CAAATTACAA AAATTCAAAA TTTTCGGGTT TATTACAGGG 
2161 ACAGCAGAGA TCCAGTTTAT CGATAAGCTT GGGAGTTCCG CGTTACATAA CTTACGGTAA 
2221 ATGGCCCGCC TGGCTGACCG CCCAACGACC CCCGCCCATT GACGTCAATA ATGACGTATG 
2281 TTCCCATAGT AACGCCAATA GGGACTTTCC ATTGACGTCA ATGGGTGGAG TATTTACGGT 
2341 AAACTGCCCA CTTGGCAGTA CATCAAGTGT ATCATATGCC AAGTACGCCC CCTATTGACG 
2401 TCAATGACGG TAAATGGCCC GCCTGGCATT ATGCCCAGTA CATGACCTTA TGGGACTTTC 
2461 CTACTTGGCA GTACATCTAC GTATTAGTCA TCGCTATTAC CATGGTGATG CGGTTTTGGC 
2521 AGTACATCAA TGGGCGTGGA TAGCGGTTTG ACTCACGGGG ATTTCCAAGT CTCCACCCCA 
2581 TTGACGTCAA TGGGAGTTTG TTTTGGCACC AAAATCAACG GGACTTTCCA AAATGTCGTA 
2641 ACAACTCCGC CCCATTGACG CAAATGGGCG GTAGGCGTGT ACGGTGGGAG GTCTATATAA 
2701 GCAGAGCTCG TTTAGTGAAC CGTCAGATCG CCTGGAGACG CCATCCACGC TGTTTTGACC 
2761 TCCATAGAAG ACACCGACTC TACTAGAGGA TCGCTAGCGC TACCGGACTC AGATCTCGAG 
2821 CTCAAGCTTC GAATTCTGCA GTCGACGGTA CCGCGGGCCC GGGATCCACC GGTCATGGTG 
2881 AGCAAGGGCG CCGAGCTGTT CACCGGCATC GTGCCCATCC TGATCGAGCT GAATGGCGAT 
2941 GTGAATGGCC ACAAGTTCAG CGTGAGCGGC GAGGGCGAGG GCGATGCCAC CTACGGCAAG 
3001 CTGACCCTGA AGTTCATCTG CACCACCGGC AAGCTGCCTG TGCCCTGGCC CACCCTGGTG 
3061 ACCACCCTGA GCTACGGCGT GCAGTGCTTC TCACGCTACC CCGATCACAT GAAGCAGCAC 
3121 GACTTCTTCA AGAGCGCCAT GCCTGAGGGC TACATCCAGG AGCGCACCAT CTTCTTCGAG 
3181 GATGACGGCA ACTACAAGTC GCGCGCCGAG GTGAAGTTCG AGGGCGATAC CCTGGTGAAT 
3241 CGCATCGAGC TGACCGGCAC CGATTTCAAG GAGGATGGCA ACATCCTGGG CAATAAGATG 
3301 GAGTACAACT ACAACGCCCA CAATGTGTAC ATCATGACCG ACAAGGCCAA GAATGGCATC 
3361 AAGGTGAACT TCAAGATCCG CCACAACATC GAGGATGGCA GCGTGCAGCT GGCCGACCAC 
3421 TACCAGCAGA ATACCCCCAT CGGCGATGGC CCTGTGCTGC TGCCCGATAA CCACTACCTG 
3481 TCCACCCAGA GCGCCCTGTC CAAGGACCCC AACGAGAAGC GCGATCACAT GATCTACTTC 
3541 GGCTTCGTGA CCGCCGCCGC CATCACCCAC GGCATGGATG AGCTGTACAA GTGAGCGGCC 
3601 GCGACTCTAG ATAATTCTAC CGGGTAGGGG AGGCGCTTTT CCCAAGGCAG TCTGGAGCAT 
3661 GCGCTTTAGC AGCCCCGCTG GGCACTTGGC GCTACACAAG TGGCCTCTGG CCTCGCACAC 
3721 ATTCCACATC CACCGGTAGG CGCCAACCGG CTCCGTTCTT TGGTGGCCCC TTCGCGCCAC 
3781 CTTCTACTCC TCCCCTAGTC AGGAAGTTCC CCCCCGCCCC GCAGCTCGCG TCGTGCAGGA 
3841 CGTGACAAAT GGAAGTAGCA CGTCTCACTA GTCTCGTGCA GATGGACAGC ACCGCTGAGC 
3901 AATGGAAGCG GGTAGGCCTT TGGGGCAGCG GCCAATAGCA GCTTTGCTCC TTCGCTTTCT 
3961 GGGCTCAGAG GCTGGGAAGG GGTGGGTCCG GGGGCGGGCT CAGGGGCGGG CTCAGGGGCG 
4021 GGGCGGGCGC CCGAAGGTCC TCCGGAGGCC CGGCATTCTG CACGCTTCAA AAGCGCACGT 
4081 CTGCCGCGCT GTTCTCCTCT TCCTCATCTC CGGGCCTTTC GACCTGCAGC CCAAGCTTAC 
4141 CATGACCGAG TACAAGCCCA CGGTGCGCCT CGCCACCCGC GACGACGTCC CCAGGGCCGT 
4201 ACGCACCCTC GCCGCCGCGT TCGCCGACTA CCCCGCCACG CGCCACACCG TCGATCCGGA 
4261 CCGCCACATC GAGCGGGTCA CCGAGCTGCA AGAACTCTTC CTCACGCGCG TCGGGCTCGA 
4321 CATCGGCAAG GTGTGGGTCG CGGACGACGG CGCCGCGGTG GCGGTCTGGA CCACGCCGGA 
4381 GAGCGTCGAA GCGGGGGCGG TGTTCGCCGA GATCGGCCCG CGCATGGCCG AGTTGAGCGG 
4441 TTCCCGGCTG GCCGCGCAGC AACAGATGGA AGGCCTCCTG GCGCCGCACC GGCCCAAGGA 
4501 GCCCGCGTGG TTCCTGGCCA CCGTCGGCGT CTCGCCCGAC CACCAGGGCA AGGGTCTGGG 
4561 CAGCGCCGTC GTGCTCCCCG GAGTGGAGGC GGCCGAGCGC GCCGGGGTGC CCGCCTTCCT 
4621 GGAGACCTCC GCGCCCCGCA ACCTCCCCTT CTACGAGCGG CTCGGCTTCA CCGTCACCGC 
4681 CGACGTCGAG GTGCCCGAAG GACCGCGCAC CTGGTGCATG ACCCGCAAGC CCGGTGCCTG 
4741 ACCGCGTCTG GAACAATCAA CCTCTGGATT ACAAAATTTG TGAAAGATTG ACTGGTATTC 
4801 TTAACTATGT TGCTCCTTTT ACGCTATGTG GATACGCTGC TTTAATGCCT TTGTATCATG 
4861 CTATTGCTTC CCGTATGGCT TTCATTTTCT CCTCCTTGTA TAAATCCTGG TTGCTGTCTC 
4921 TTTATGAGGA GTTGTGGCCC GTTGTCAGGC AACGTGGCGT GGTGTGCACT GTGTTTGCTG 
4981 ACGCAACCCC CACTGGTTGG GGCATTGCCA CCACCTGTCA GCTCCTTTCC GGGACTTTCG 
5041 CTTTCCCCCT CCCTATTGCC ACGGCGGAAC TCATCGCCGC CTGCCTTGCC CGCTGCTGGA 
5101 CAGGGGCTCG GCTGTTGGGC ACTGACAATT CCGTGGTGTT GTCGGGGAAG CTGACGTCCT 
5161 TTCCATGGCT GCTCGCCTGT GTTGCCACCT GGATTCTGCG CGGGACGTCC TTCTGCTACG 
5221 TCCCTTCGGC CCTCAATCCA GCGGACCTTC CTTCCCGCGG CCTGCTGCCG GCTCTGCGGC 
5281 CTCTTCCGCG TCTTCGCCTT CGCCCTCAGA CGAGTCGGAT CTCCCTTTGG GCCGCCTCCC 
5341 CGCCTGGAAT TAATTCTGCA GTCGAGACCT AGAAAAACAT GGAGCAATCA CAAGTAGCAA 
5401 TACAGCAGCT ACCAATGCTG ATTGTGCCTG GCTAGAAGCA CAAGAGGAGG AGGAGGTGGG 
5461 TTTTCCAGTC ACACCTCAGG TACCTTTAAG ACCAATGACT TACAAGGCAG CTGTAGATCT 
5521 TAGCCACTTT TTAAAAGAAA AGAGGGGACT GGAAGGGCTA ATTCACTCCC AACGAAGACA 
5581 AGATATCCTT GATCTGTGGA TCTACCACAC ACAAGGCTAC TTCCCTGATT AGCAGAACTA 
5641 CACACCAGGG CCAGGGGTCA GATATCCACT GACCTTTGGA TGGTGCTACA AGCTAGTACC 
5701 AGTTGAGCCA GATAAGGTAG AAGAGGCCAA TAAAGGAGAG AACACCAGCT TGTTACACCC 
5761 TGTGAGCCTG CATGGGATGG ATGACCCGGA GAGAGAAGTG TTAGAGTGGA GGTTTGACAG 
5821 CCGCCTAGCA TTTCATCACG TGGCCCGAGA GCTGCATCCG GAGTACTTCA AGAACTGCTG 
5881 ATATCGAGCT TGCTACAAGG GACTTTCCGC TGGGGACTTT CCAGGGAGGC GTGGCCTGGG 
5941 CGGGACTGGG GAGTGGCGAG CCCTCAGATC CTGCATATAA GCAGCTGCTT TTTGCCTGTA 
6001 CTGGGTCTCT CTGGTTAGAC CAGATCTGAG CCTGGGAGCT CTCTGGCTAA CTAGGGAACC 
6061 CACTGCTTAA GCCTCAATAA AGCTTGCCTT GAGTGCTTCA AGTAGTGTGT GCCCGTCTGT 
6121 TGTGTGACTC TGGTAACTAG AGATCCCTCA GACCCTTTTA GTCAGTGTGG AAAATCTCTA 
6181 GCAGTAGTAG TTCATGTCAT CTTATTATTC AGTATTTATA ACTTGCAAAG AAATGAATAT 
6241 CAGAGAGTGA GAGGCCTTGA CATTGCTAGC GTTTTACCGT CGACCTCTAG CTAGAGCTTG 
6301 GCGTAATCAT GGTCATAGCT GTTTCCTGTG TGAAATTGTT ATCCGCTCAC AATTCCACAC 
6361 AACATACGAG CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG CCTAATGAGT GAGCTAACTC 
6421 ACATTAATTG CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG GAAACCTGTC GTGCCAGCTG 
6481 CATTAATGAA TCGGCCAACG CGCGGGGAGA GGCGGTTTGC GTATTGGGCG CTCTTCCGCT 
6541 TCCTCGCTCA CTGACTCGCT GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT ATCAGCTCAC 
6601 TCAAAGGCGG TAATACGGTT ATCCACAGAA TCAGGGGATA ACGCAGGAAA GAACATGTGA 
6661 GCAAAAGGCC AGCAAAAGGC CAGGAACCGT AAAAAGGCCG CGTTGCTGGC GTTTTTCCAT 
6721 AGGCTCCGCC CCCCTGACGA GCATCACAAA AATCGACGCT CAAGTCAGAG GTGGCGAAAC 
6781 CCGACAGGAC TATAAAGATA CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT GCGCTCTCCT 
6841 GTTCCGACCC TGCCGCTTAC CGGATACCTG TCCGCCTTTC TCCCTTCGGG AAGCGTGGCG 
6901 CTTTCTCATA GCTCACGCTG TAGGTATCTC AGTTCGGTGT AGGTCGTTCG CTCCAAGCTG 
6961 GGCTGTGTGC ACGAACCCCC CGTTCAGCCC GACCGCTGCG CCTTATCCGG TAACTATCGT 
7021 CTTGAGTCCA ACCCGGTAAG ACACGACTTA TCGCCACTGG CAGCAGCCAC TGGTAACAGG 
7081 ATTAGCAGAG CGAGGTATGT AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG GCCTAACTAC 
7141 GGCTACACTA GAAGAACAGT ATTTGGTATC TGCGCTCTGC TGAAGCCAGT TACCTTCGGA 
7201 AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA CAAACCACCG CTGGTAGCGG TGGTTTTTTT 
7261 GTTTGCAAGC AGCAGATTAC GCGCAGAAAA AAAGGATCTC AAGAAGATCC TTTGATCTTT 
7321 TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT GGTCATGAGA 
7381 TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA AATGAAGTTT TAAATCAATC 
7441 TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT GCTTAATCAG TGAGGCACCT 
7501 ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT GACTCCCCGT CGTGTAGATA 
7561 ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG CAATGATACC GCGAGACCCA 
7621 CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG CCGGAAGGGC CGAGCGCAGA 
7681 AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA ATTGTTGCCG GGAAGCTAGA 
7741 GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG CCATTGCTAC AGGCATCGTG 
7801 GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG GTTCCCAACG ATCAAGGCGA 
7861 GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC TCCGATCGTT 
7921 GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA TGGCAGCACT GCATAATTCT 
7981 CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG GTGAGTACTC AACCAAGTCA 
8041 TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC CGGCGTCAAT ACGGGATAAT 
8101 ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG GAAAACGTTC TTCGGGGCGA 
8161 AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA TGTAACCCAC TCGTGCACCC 
8221 AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG GGTGAGCAAA AACAGGAAGG 
8281 CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT GTTGAATACT CATACTCTTC 
8341 CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC TCATGAGCGG ATACATATTT 
8401 GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA CATTTCCCCG AAAAGTGCCA 
8461 CCTGACGTCG ACGGATCGGG AGATCAACTT GTTTATTGCA GCTTATAATG GTTACAAATA 
8521 AAGCAATAGC ATCACAAATT TCACAAATAA AGCATTTTTT TCACTGCATT CTAGTTGTGG 
8581 TTTGTCCAAA CTCATCAATG TATCTTATCA TGTCTGGATC AACTGGATAA CTCAAGCTAA 
8641 CCAAAATCAT CCCAAACTTC CCACCCCATA CCCTATTACC ACTGCCAATT ACCTGTGGTT 
8701 TCATTTACTC TAAACCTGTG ATTCCTCTGA ATTATTTTCA TTTTAAAGAA ATTGTATTTG 
8761 TTAAATATGT ACTACAAACT TAGTAGT