pLPCX

pLPCX contains elements derived from Moloney murine leukemia virus (MoMuLV) and Moloney murine sarcoma virus (MoMuSV), and is designed for retroviral gene delivery and expression (1–3). Upon transfection into a packaging cell line, pLPCX transiently expresses, or integrates and stably expresses, a transcript containing Ψ+ (the extended viral packaging signal) the puromycin resistance gene, and the gene of interest. The 5' viral LTR in this vector contains promoter/enhancer sequences that control expression of the puromycin resistance (Puror) gene for antibiotic selection in eukaryotic cells. A gene of interest can be cloned into the multiple cloning site immediately downstream of the human cytomegalovirus (CMV) immediate early promoter (PCMV IE). pLPCX also includes the Col E1 origin of replication and E. coli Ampr gene for propagation and antibiotic selection in bacteria.

载体应用

pLPCX can be transfected into a packaging cell line such as the RetroPackTM PT67 Cell Line (Cat. No. 631510). Once in the cell, RNA from the vector is packaged into infectious, replication- incompetent retroviral particles. pLPCX does not contain the structural genes (gag, pol, and env) necessary for particle formation and replication; however, these genes are stably integrated into PT67 (4–7). Subsequent introduction of pLPCX, containing Ψ+, transcription and processing elements, and the gene of interest produces high-titer, replication-incompetent infectious virus. These retroviral particles can infect target cells and transmit the gene of interest (which is cloned between the viral LTR sequences), but cannot replicate within these cells since the cells lack the viral structural genes. The separate introduction and integration of the structural genes into the packaging cell line minimizes the chances of producing replication- competent virus due to recombination events during cell proliferation.

0001 TTTGAAAGAC CCCACCCGTA GGTGGCAAGC TAGCTTAAGT AACGCCACTT TGCAAGGCAT 
0061 GGAAAAATAC ATAACTGAGA ATAGAAAAGT TCAGATCAAG GTCAGGAACA AAGAAACAGC 
0121 TGAATACCAA ACAGGATATC TGTGGTAAGC GGTTCCTGCC CCGGCTCAGG GCCAAGAACA 
0181 GATGAGACAG CTGAGTGATG GGCCAAACAG GATATCTGTG GTAAGCAGTT CCTGCCCCGG 
0241 CTCGGGGCCA AGAACAGATG GTCCCCAGAT GCGGTCCAGC CCTCAGCAGT TTCTAGTGAA 
0301 TCATCAGATG TTTCCAGGGT GCCCCAAGGA CCTGAAAATG ACCCTGTACC TTATTTGAAC 
0361 TAACCAATCA GTTCGCTTCT CGCTTCTGTT CGCGCGCTTC CGCTCTCCGA GCTCAATAAA 
0421 AGAGCCCACA ACCCCTCACT CGGCGCGCCA GTCTTCCGAT AGACTGCGTC GCCCGGGTAC 
0481 CCGTATTCCC AATAAAGCCT CTTGCTGTTT GCATCCGAAT CGTGGTCTCG CTGTTCCTTG 
0541 GGAGGGTCTC CTCTGAGTGA TTGACTACCC ACGACGGGGG TCTTTCATTT GGGGGCTCGT 
0601 CCGGGATTTG GAGACCCCTG CCCAGGGACC ACCGACCCAC CACCGGGAGG TAAGCTGGCC 
0661 AGCAACTTAT CTGTGTCTGT CCGATTGTCT AGTGTCTATG TTTGATGTTA TGCGCCTGCG 
0721 TCTGTACTAG TTAGCTAACT AGCTCTGTAT CTGGCGGACC CGTGGTGGAA CTGACGAGTT 
0781 CTGAACACCC GGCCGCAACC CTGGGAGACG TCCCAGGGAC TTTGGGGGCC GTTTTTGTGG 
0841 CCCGACCTGA GGAAGGGAGT CGATGTGGAA TCCGACCCCG TCAGGATATG TGGTTCTGGT 
0901 AGGAGACGAG AACCTAAAAC AGTTCCCGCC TCCGTCTGAA TTTTTGCTTT CGGTTTGGAA 
0961 CCGAAGCCGC GCGTCTTGTC TGCTGCAGCG CTGCAGCATC GTTCTGTGTT GTCTCTGTCT 
1021 GACTGTGTTT CTGTATTTGT CTGAAAATTA GGGCCAGACT GTTACCACTC CCTTAAGTTT 
1081 GACCTTAGGT CACTGGAAAG ATGTCGAGCG GATCGCTCAC AACCAGTCGG TAGATGTCAA 
1141 GAAGAGACGT TGGGTTACCT TCTGCTCTGC AGAATGGCCA ACCTTTAACG TCGGATGGCC 
1201 GCGAGACGGC ACCTTTAACC GAGACCTCAT CACCCAGGTT AAGATCAAGG TCTTTTCACC 
1261 TGGCCCGCAT GGACACCCAG ACCAGGTCCC CTACATCGTG ACCTGGGAAG CCTTGGCTTT 
1321 TGACCCCCCT CCCTGGGTCA AGCCCTTTGT ACACCCTAAG CCTCCGCCTC CTCTTCCTCC 
1381 ATCCGCCCCG TCTCTCCCCC TTGAACCTCC TCGTTCGACC CCGCCTCGAT CCTCCCTTTA 
1441 TCCAGCCCTC ACTCCTTCTC TAGGCGCCGG AATTGATCCC CCGGGCTGCA GGTCGGCCGC 
1501 CACGACCGGT GCCGCCACCA TCCCCTGACC CACGCCCCTG ACCCCTCACA AGGAGACGAC 
1561 CTTCCATGAC CGAGTACAAG CCCACGGTGC GCCTCGCCAC CCGCGACGAC GTCCCCCGGG 
1621 CCGTACGCAC CCTCGCCGCC GCGTTCGCCG ACTACCCCGC CACGCGCCAC ACCGTCGACC 
1681 CGGACCGCCA CATCGAGCGG GTCACCGAGC TGCAAGAACT CTTCCTCACG CGCGTCGGGC 
1741 TCGACATCGG CAAGGTGTGG GTCGCGGACG ACGGCGCCGC GGTGGCGGTC TGGACCACGC 
1801 CGGAGAGCGT CGAAGCGGGG GCGGTGTTCG CCGAGATCGG CCCGCGCATG GCCGAGTTGA 
1861 GCGGTTCCCG GCTGGCCGCG CAGCAACAGA TGGAAGGCCT CCTGGCGCCG CACCGGCCCA 
1921 AGGAGCCCGC GTGGTTCCTG GCCACCGTCG GCGTCTCGCC CGACCACCAG GGCAAGGGTC 
1981 TGGGCAGCGC CGTCGTGCTC CCCGGAGTGG AGGCGGCCGA GCGCGCCGGG GTGCCCGCCT 
2041 TCCTGGAGAC CTCCGCGCCC CGCAACCTCC CCTTCTACGA GCGGCTCGGC TTCACCGTCA 
2101 CCGCCGACGT CGAGTGCCCG AAGGACCGCG CGACCTGGTG CATGACCCGC AAGCCCGGTG 
2161 CCTGACGCCC GCCCCACGAC CCGCAGCGCC CGACCGAAAG GAGCGCACGA CCCCATGGCT 
2221 CCGACCGAAG CCGACCCGGG CGGCCCCGCC GACCCCGCAC CCGCCCCCGA GGCCCACCGA 
2281 CTCTAGTAAT AGTAATCAAT TACGGGGTCA TTAGTTCATA GCCCATATAT GGAGTTCCGC 
2341 GTTACATAAC TTACGGTAAA TGGCCCGCCT GGCTGACCGC CCAACGACCC CCGCCCATTG 
2401 ACGTCAATAA TGACGTATGT TCCCATAGTA ACGCCAATAG GGACTTTCCA TTGACGTCAA 
2461 TGGGTGGAGT ATTTACGGTA AACTGCCCAC TTGGCAGTAC ATCAAGTGTA TCATATGCCA 
2521 AGTACGCCCC CTATTGACGT CAATGACGGT AAATGGCCCG CCTGGCATTA TGCCCAGTAC 
2581 ATGACCTTAT GGGACTTTCC TACTTGGCAG TACATCTACG TATTAGTCAT CGCTATTACC 
2641 ATGGTGATGC GGTTTTGGCA GTACATCAAT GGGCGTGGAT AGCGGTTTGA CTCACGGGGA 
2701 TTTCCAAGTC TCCACCCCAT TGACGTCAAT GGGAGTTTGT TTTGGCACCA AAATCAACGG 
2761 GACTTTCCAA AATGTCGTAA CAACTCCGCC CCATTGACGC AAATGGGCGG TAGGCGTGTA 
2821 CGGTGGGAGG TCTATATAAG CAGAGCTGGT TTAGTGAACC GTCAGATCCG CTAGCGCTAC 
2881 CGGACTCAGA TCTCGAGCTC AAGCTTCGAA TTCGTTAGGC CATTAAGGCC TGTCGACAAG 
2941 CGGCCGCCTC GGCCAAACAT CGATAAAATA AAAGATTTTA TTTAGTCTCC AGAAAAAGGG 
3001 GGGAATGAAA GACCCCACCT GTAGGTTTGG CAAGCTAGCT TAAGTAACGC CATTTTGCAA 
3061 GGCATGGAAA AATACATAAC TGAGAATAGA GAAGTTCAGA TCAAGGTCAG GAACAGATGG 
3121 AACAGCTGAA TATGGGCCAA ACAGGATATC TGTGGTAAGC AGTTCCTGCC CCGGCTCAGG 
3181 GCCAAGAACA GATGGAACAG CTGAATATGG GCCAAACAGG ATATCTGTGG TAAGCAGTTC 
3241 CTGCCCCGGC TCAGGGCCAA GAACAGATGG TCCCCAGATG CGGTCCAGCC CTCAGCAGTT 
3301 TCTAGAGAAC CATCAGATGT TTCCAGGGTG CCCCAAGGAC CTGAAATGAC CCTGTGCCTT 
3361 ATTTGAACTA ACCAATCAGT TCGCTTCTCG CTTCTGTTCG CGCGCTTCTG CTCCCCGAGC 
3421 TCAATAAAAG AGCCCACAAC CCCTCACTCG GGGCGCCAGT CCTCCGATTG ACTGAGTCGC 
3481 CCGGGTACCC GTGTATCCAA TAAACCCTCT TGCAGTTGCA TCCGACTTGT GGTCTCGCTG 
3541 TTCCTTGGGA GGGTCTCCTC TGAGTGATTG ACTACCCGTC AGCGGGGGTC TTTCATTTGG 
3601 GGGCTCGTCC GGGATCGGGA GACCCCTGCC CAGGGACCAC CGACCCACCA CCGGGAGGTA 
3661 AGCTGGCTGC CTCGCGCGTT TCGGTGATGA CGGTGAAAAC CTCTGACACA TGCAGCTCCC 
3721 GGAGACGGTC ACAGCTTGTC TGTAAGCGGA TGCCGGGAGC AGACAAGCCC GTCAGGGCGC 
3781 GTCAGCGGGT GTTGGCGGGT GTCGGGGCGC AGCCATGACC CAGTCACGTA GCGATAGCGG 
3841 AGTGTACTGA TAACTTCGTA TAATGTATGC TATACGAAGT TATTAGGTCT GAAGAGGAGT 
3901 TTACGTCCAG CCAAGCTAGG GCCGCGATCC GGAACCCTTA ATATAACTTC GTATAATGTA 
3961 TGCTATACGA AGTTATCAGT ACTGGCTTAA CTATGCGGCA TCAGAGCAGA TTGTACTGAG 
4021 AGTGCACCAT ATGCGGTGTG AAATACCGCA CAGATGCGTA AGGAGAAAAT ACCGCATCAG 
4081 GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC 
4141 GGTATCAGCT CACTCAAAGG CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGG 
4201 AAAGAACATG TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT 
4261 GGCGTTTTTC CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA 
4321 GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT 
4381 CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC 
4441 GGGAAGCGTG GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT 
4501 TCGCTCCAAG CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC 
4561 CGGTAACTAT CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC 
4621 CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT TCTTGAAGTG 
4681 GTGGCCTAAC TACGGCTACA CTAGAAGGAC AGTATTTGGT ATCTGCGCTC TGCTGAAGCC 
4741 AGTTACCTTC GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA CCGCTGGTAG 
4801 CGGTGGTTTT TTTGTTTGCA AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA 
4861 TCCTTTGATC TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT 
4921 TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT AAAAATGAAG 
4981 TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT GACAGTTACC AATGCTTAAT 
5041 CAGTGAGGCA CCTATCTCAG CGATCTGTCT ATTTCGTTCA TCCATAGTTG CCTGACTCCC 
5101 CGTCGTGTAG ATAACTACGA TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT 
5161 ACCGCGAGAC CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG 
5221 GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA TTAATTGTTG 
5281 CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG CGCAACGTTG TTGCCATTGC 
5341 TGCAGGCATC GTGGTGTCAC GCTCGTCGTT TGGTATGGCT TCATTCAGCT CCGGTTCCCA 
5401 ACGATCAAGG CGAGTTACAT GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG 
5461 TCCTCCGATC GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC 
5521 ACTGCATAAT TCTCTTACTG TCATGCCATC CGTAAGATGC TTTTCTGTGA CTGGTGAGTA 
5581 CTCAACCAAG TCATTCTGAG AATAGTGTAT GCGGCGACCG AGTTGCTCTT GCCCGGCGTC 
5641 AACACGGGAT AATACCGCGC CACATAGCAG AACTTTAAAA GTGCTCATCA TTGGAAAACG 
5701 TTCTTCGGGG CGAAAACTCT CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC 
5761 CACTCGTGCA CCCAACTGAT CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC 
5821 AAAAACAGGA AGGCAAAATG CCGCAAAAAA GGGAATAAGG GCGACACGGA AATGTTGAAT 
5881 ACTCATACTC TTCCTTTTTC AATATTATTG AAGCATTTAT CAGGGTTATT GTCTCATGAG 
5941 CGGATACATA TTTGAATGTA TTTAGAAAAA TAAACAAATA GGGGTTCCGC GCACATTTCC 
6001 CCGAAAAGTG CCACCTGACG TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA 
6061 TAGGCGTATC ACGAGGCCCT TTCGTCTTCA AGAATTGCTA GCAATTGCTA GCAATTGCTA 
6121 GCAATTCATA CCAGATCACC GAAAACTGTC CTCCAAATGT GTCCCCCTCA CACTCCCAAA 
6181 TTCGCGGGCT TCTGCCTCTT AGACCACTCT ACCCTATTCC CCACACTCAC CGGAGCCAAA 
6241 GCCGCGGCCC TTCCGTTTCT TTGCT