测序及更多信息：https://pan.baidu.com/s/1tCmowAl5tOi_BQMKk4UGBQ
提取码：868m
采购地址：淘宝店铺地址
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EBNA1 和oriP 是EB 病毒中与复制和保持病毒基因组游离性有关的重要基因元件。EBNA1 为EB 病毒核抗原1 的基因, 编码一个磷酸化的核蛋白, 蛋白的C 端为一个二聚化的DNA 结合区, EBNA1 同源二聚化与DNA 上的特异序列结合。Or iP 含有双对称( dyad symmetry DS) 和成簇的重复序列( the family of repeats FR) 。这些序列均包含与EBNA1 结合的多个同源序列, DS 区含有约共120bp 的四个EBNA1 结合位点, FR 区包含有大约20 个串联重复的30bp 序列。当EBNA1与这些序列结合后, 使病毒的基因组以稳定的游离体形式存在, 并使病毒基因组依赖宿主复制的方式复制。其复制与宿主细胞染色体复制的前S 期同步, 一个细胞周期只复制一次。游离体在分裂过程中, 通过EBNA1 以非共价的方式与宿主染色体结合, 保证了分裂时游离体进行正确的分离 。除与复制有关外, EBNA1 还有利于游离基因组的核转移, 核定位信号位于EBNA1 蛋白的中部, 其作用与转运蛋白karyopherinA2 相关 。EBNA1 也能与RNA 结合, 对基因表达进行转录后调节[3] 。此外, EBNA1 介导EB 病毒对宿主的免疫逃逸 。OriP 序列的FR 区是依赖于EBNA1 的转录增强子。
鉴于EBNA1PoriP 的特点, Yate 在1985 年首次将这两个基因元件引入质粒。具有EBNA1PoriP 的质粒转染入靶细胞后不整合, 可以长期稳定存在, 不会造成宿主的突变 。同时, 由于它的转运入核、转录增强、免疫逃逸等功能, 使质粒携带的目的基因能够获得较高的转染效率, 高水平、持续的表达。因此, 基于EBNA1PoriP 的质粒是非病毒型基因治疗载体中比较理想的一个, 在多个基因治疗方案中, 显示了良好的应用前景。
参考文献
An Efficient Non-viral Method to Generate Integration-Free Human iPS Cells from Cord Blood and Peripheral Blood Cells. Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A, Goshima N, Yamanaka S. Stem Cells. 2012 Nov 29. doi: 10.1002/stem.1293. 10.1002/stem.1293PubMed 23193063
全序列
1 TCGACATTGA TTATTGACTA GTTATTAATA GTAATCAATT ACGGGGTCAT TAGTTCATAG
61 CCCATATATG GAGTTCCGCG TTACATAACT TACGGTAAAT GGCCCGCCTG GCTGACCGCC
121 CAACGACCCC CGCCCATTGA CGTCAATAAT GACGTATGTT CCCATAGTAA CGCCAATAGG
181 GACTTTCCAT TGACGTCAAT GGGTGGAGTA TTTACGGTAA ACTGCCCACT TGGCAGTACA
241 TCAAGTGTAT CATATGCCAA GTACGCCCCC TATTGACGTC AATGACGGTA AATGGCCCGC
301 CTGGCATTAT GCCCAGTACA TGACCTTATG GGACTTTCCT ACTTGGCAGT ACATCTACGT
361 ATTAGTCATC GCTATTACCA TGGTCGAGGT GAGCCCCACG TTCTGCTTCA CTCTCCCCAT
421 CTCCCCCCCC TCCCCACCCC CAATTTTGTA TTTATTTATT TTTTAATTAT TTTGTGCAGC
481 GATGGGGGCG GGGGGGGGGG GGGGGCGCGC GCCAGGCGGG GCGGGGCGGG GCGAGGGGCG
541 GGGCGGGCGA GGCGGAGAGG TGCGGCGGCA GCCAATCAGA GCGGCGCGCT CCGAAAGTTT
601 CCTTTTATGG CGAGGCGGCG GCGGCGGCGG CCCTATAAAA AGCGAAGCGC GCGGCGGGCG
661 GGAGTCGCTG CGCGCTGCCT TCGCCCCGTG CCCCGCTCCG CCGCCGCCTC GCGCCGCCCG
721 CCCCGGCTCT GACTGACCGC GTTACTCCCA CAGGTGAGCG GGCGGGACGG CCCTTCTCCT
781 CCGGGCTGTA ATTAGCGCTT GGTTTAATGA CGGCTTGTTT CTTTTCTGTG GCTGCGTGAA
841 AGCCTTGAGG GGCTCCGGGA GGGCCCTTTG TGCGGGGGGA GCGGCTCGGG GGGTGCGTGC
901 GTGTGTGTGT GCGTGGGGAG CGCCGCGTGC GGCTCCGCGC TGCCCGGCGG CTGTGAGCGC
961 TGCGGGCGCG GCGCGGGGCT TTGTGCGCTC CGCAGTGTGC GCGAGGGGAG CGCGGCCGGG
1021 GGCGGTGCCC CGCGGTGCGG GGGGGGCTGC GAGGGGAACA AAGGCTGCGT GCGGGGTGTG
1081 TGCGTGGGGG GGTGAGCAGG GGGTGTGGGC GCGTCGGTCG GGCTGCAACC CCCCCTGCAC
1141 CCCCCTCCCC GAGTTGCTGA GCACGGCCCG GCTTCGGGTG CGGGGCTCCG TACGGGGCGT
1201 GGCGCGGGGC TCGCCGTGCC GGGCGGGGGG TGGCGGCAGG TGGGGGTGCC GGGCGGGGCG
1261 GGGCCGCCTC GGGCCGGGGA GGGCTCGGGG GAGGGGCGCG GCGGCCCCCG GAGCGCCGGC
1321 GGCTGTCGAG GCGCGGCGAG CCGCAGCCAT TGCCTTTTAT GGTAATCGTG CGAGAGGGCG
1381 CAGGGACTTC CTTTGTCCCA AATCTGTGCG GAGCCGAAAT CTGGGAGGCG CCGCCGCACC
1441 CCCTCTAGCG GGCGCGGGGC GAAGCGGTGC GGCGCCGGCA GGAAGGAAAT GGGCGGGGAG
1501 GGCCTTCGTG CGTCGCCGCG CCGCCGTCCC CTTCTCCCTC TCCAGCCTCG GGGCTGTCCG
1561 CGGGGGGACG GCTGCCTTCG GGGGGGACGG GGCAGGGCGG GGTTCGGCTT CTGGCGTGTG
1621 ACCGGCGGCT CTAGAGCCTC TGCTAACCAT GTTCATGCCT TCTTCTTTTT CCTACAGCTC
1681 CTGGGCAACG TGCTGGTTAT TGTGCTGTCT CATCATTTTG GCAAAGAATT CGCCCTTGTC
1741 GACACCATGT CTGACGAGGG GCCAGGTACA GGACCTGGAA ATGGCCTAGG AGAGAAGGGA
1801 GACACATCTG GACCAGAAGG CTCCGGCGGC AGTGGACCTC AAAGAAGAGG GGGTGATAAC
1861 CATGGACGAG GACGGGGAAG AGGACGAGGA CGAGGAGGCG GAAGACCAGG AGCCCCGGGC
1921 GGCTCAGGAT CAGGGCCAAG ACATAGAGAT GGTGTCCGGA GACCCCAAAA ACGTCCAAGT
1981 TGCATTGGCT GCAAAGGGAC CCACGGTGGA ACAGGAGCAG GAGCAGGAGC GGGAGGGGCA
2041 GGAGCAGGAG GGGCAGGAGC AGGAGGAGGG GCAGGAGCAG GAGGAGGGGC AGGAGGGGCA
2101 GGAGGGGCAG GAGGGGCAGG AGCAGGAGGA GGGGCAGGAG CAGGAGGAGG GGCAGGAGGG
2161 GCAGGAGGGG CAGGAGCAGG AGGAGGGGCA GGAGCAGGAG GAGGGGCAGG AGGGGCAGGA
2221 GCAGGAGGAG GGGCAGGAGG GGCAGGAGGG GCAGGAGCAG GAGGAGGGGC AGGAGCAGGA
2281 GGAGGGGCAG GAGGGGCAGG AGCAGGAGGA GGGGCAGGAG GGGCAGGAGG GGCAGGAGCA
2341 GGAGGAGGGG CAGGAGCAGG AGGGGCAGGA GGGGCAGGAG GGGCAGGAGC AGGAGGGGCA
2401 GGAGCAGGAG GAGGGGCAGG AGGGGCAGGA GGGGCAGGAG CAGGAGGGGC AGGAGCAGGA
2461 GGGGCAGGAG CAGGAGGGGC AGGAGCAGGA GGGGCAGGAG GGGCAGGAGC AGGAGGGGCA
2521 GGAGGGGCAG GAGCAGGAGG GGCAGGAGGG GCAGGAGCAG GAGGAGGGGC AGGAGGGGCA
2581 GGAGCAGGAG GAGGGGCAGG AGGGGCAGGA GCAGGAGGGG CAGGAGGGGC AGGAGCAGGA
2641 GGGGCAGGAG GGGCAGGAGC AGGAGGGGCA GGAGGGGCAG GAGCAGGAGG AGGGGCAGGA
2701 GCAGGAGGGG CAGGAGCAGG AGGTGGAGGC CGGGGTCGAG GAGGCAGTGG AGGCCGGGGT
2761 CGAGGAGGTA GTGGAGGCCG GGGTCGAGGA GGTAGTGGAG GCCGCCGGGG TAGAGGACGT
2821 GAAAGAGCCA GGGGGGGAAG TCGTGAAAGA GCCAGGGGGA GAGGTCGTGG ACGTGGAGAA
2881 AAGAGGCCCA GGAGTCCCAG TAGTCAGTCA TCATCATCCG GGTCTCCACC GCGCAGGCCC
2941 CCTCCAGGTA GAAGGCCATT TTTCCACCCT GTAGGGGAAG CCGATTATTT TGAATACCAC
3001 CAAGAAGGTG GCCCAGATGG TGAGCCTGAC GTGCCCCCGG GAGCGATAGA GCAGGGCCCC
3061 GCAGATGACC CAGGAGAAGG CCCAAGCACT GGACCCCGGG GTCAGGGTGA TGGAGGCAGG
3121 CGCAAAAAAG GAGGGTGGTT TGGAAAGCAT CGTGGTCAAG GAGGTTCCAA CCCGAAATTT
3181 GAGAACATTG CAGAAGGTTT AAGAGCTCTC CTGGCTAGGA GTCACGTAGA AAGGACTACC
3241 GACGAAGGAA CTTGGGTCGC CGGTGTGTTC GTATATGGAG GTAGTAAGAC CTCCCTTTAC
3301 AACCTAAGGC GAGGAACTGC CCTTGCTATT CCACAATGTC GTCTTACACC ATTGAGTCGT
3361 CTCCCCTTTG GAATGGCCCC TGGACCCGGC CCACAACCTG GCCCGCTAAG GGAGTCCATT
3421 GTCTGTTATT TCATGGTCTT TTTACAAACT CATATATTTG CTGAGGTTTT GAAGGATGCG
3481 ATTAAGGACC TTGTTATGAC AAAGCCCGCT CCTACCTGCA ATATCAGGGT GACTGTGTGC
3541 AGCTTTGACG ATGGAGTAGA TTTGCCTCCC TGGTTTCCAC CTATGGTGGA AGGGGCTGCC
3601 GCGGAGGGTG ATGACGGAGA TGACGGAGAT GAAGGAGGTG ATGGAGATGA GGGTGAGGAA
3661 GGGCAGGAGT GAGGATCCAA GGGCGAATTC AAGCTTCGGG GACTAGTCAT ATGATAATCA
3721 ACCTCTGGAT TACAAAATTT GTGAAAGATT GACTGGTATT CTTAACTATG TTGCTCCTTT
3781 TACGCTATGT GGATACGCTG CTTTAATGCC TTTGTATCAT GCTATTGCTT CCCGTATGGC
3841 TTTCATTTTC TCCTCCTTGT ATAAATCCTG GTTGCTGTCT CTTTATGAGG AGTTGTGGCC
3901 CGTTGTCAGG CAACGTGGCG TGGTGTGCAC TGTGTTTGCT GACGCAACCC CCACTGGTTG
3961 GGGCATTGCC ACCACCTGTC AGCTCCTTTC CGGGACTTTC GCTTTCCCCC TCCCTATTGC
4021 CACGGCGGAA CTCATCGCCG CCTGCCTTGC CCGCTGCTGG ACAGGGGCTC GGCTGTTGGG
4081 CACTGACAAT TCCGTGGTGT TGTCGGGGAA GCTGACGTCC TTTCCATGGC TGCTCGCCTG
4141 TGTTGCCACC TGGATTCTGC GCGGGACGTC CTTCTGCTAC GTCCCTTCGG CCCTCAATCC
4201 AGCGGACCTT CCTTCCCGCG GCCTGCTGCC GGCTCTGCGG CCTCTTCCGC GTCTTCGCCT
4261 TCGCCCTCAG ACGAGTCGGA TCTCCCTTTG GGCCGCCTCC CCGCATCGGT AAATTCACTC
4321 CTCAGGTGCA GGCTGCCTAT CAGAAGGTGG TGGCTGGTGT GGCCAATGCC CTGGCTCACA
4381 AATACCACTG AGATCTTTTT CCCTCTGCCA AAAATTATGG GGACATCATG AAGCCCCTTG
4441 AGCATCTGAC TTCTGGCTAA TAAAGGAAAT TTATTTTCAT TGCAATAGTG TGTTGGAATT
4501 TTTTGTGTCT CTCACTCGGA AGGACATATG GGAGGGCAAA TCATTTAAAA CATCAGAATG
4561 AGTATTTGGT TTAGAGTTTG GCAACATATG CCCATATGCT GGCTGCCATG AACAAAGGTT
4621 GGCTATAAAG AGGTCATCAG TATATGAAAC AGCCCCCTGC TGTCCATTCC TTATTCCATA
4681 GAAAAGCCTT GACTTGAGGT TAGATTTTTT TTATATTTTG TTTTGTGTTA TTTTTTTCTT
4741 TAACATCCCT AAAATTTTCC TTACATGTTT TACTAGCCAG ATTTTTCCTC CTCTCCTGAC
4801 TACTCCCAGT CATAGCTGTC CCTCTTCTCT TATGGAGATC CCTCGACCTG CAGCCCAAGC
4861 TTGGCGTAAT CATGGTCATA GCTGTTTCCT GTGTGAAATT GTTATCCGCT CACAATTCCA
4921 CACAACATAC GAGCCGGAAG CATAAAGTGT AAAGCCTGGG GTGCCTAATG AGTGAGCTAA
4981 CTCACATTAA TTGCGTTGCG CTCACTGCCC GCTTTCCAGT CGGGAAACCT GTCGTGCCAG
5041 CGGATCCGCT GCATTAATGA ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC
5101 GCTCTTCCGC TTCCTCGCTC ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG
5161 TATCAGCTCA CTCAAAGGCG GTAATACGGT TATCCACAGA ATCAGGGGAT AACGCAGGAA
5221 AGAACATGTG AGCAAAAGGC CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG
5281 CGTTTTTCCA TAGGCTCCGC CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA
5341 GGTGGCGAAA CCCGACAGGA CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG
5401 TGCGCTCTCC TGTTCCGACC CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG
5461 GAAGCGTGGC GCTTTCTCAT AGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC
5521 GCTCCAAGCT GGGCTGTGTG CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG
5581 GTAACTATCG TCTTGAGTCC AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA
5641 CTGGTAACAG GATTAGCAGA GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT
5701 GGCCTAACTA CGGCTACACT AGAAGAACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG
5761 TTACCTTCGG AAAAAGAGTT GGTAGCTCTT GATCCGGCAA ACAAACCACC GCTGGTAGCG
5821 GTGGTTTTTT TGTTTGCAAG CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC
5881 CTTTGATCTT TTCTACGGGG TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT
5941 TGGTCATGAG ATTATCAAAA AGGATCTTCA CCTAGATCCT TTTAAATTAA AAATGAAGTT
6001 TTAAATCAAT CTAAAGTATA TATGAGTAAA CTTGGTCTGA CAGTTACCAA TGCTTAATCA
6061 GTGAGGCACC TATCTCAGCG ATCTGTCTAT TTCGTTCATC CATAGTTGCC TGACTCCCCG
6121 TCGTGTAGAT AACTACGATA CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC
6181 CGCGAGACCC ACGCTCACCG GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG
6241 CCGAGCGCAG AAGTGGTCCT GCAACTTTAT CCGCCTCCAT CCAGTCTATT AATTGTTGCC
6301 GGGAAGCTAG AGTAAGTAGT TCGCCAGTTA ATAGTTTGCG CAACGTTGTT GCCATTGCTA
6361 CAGGCATCGT GGTGTCACGC TCGTCGTTTG GTATGGCTTC ATTCAGCTCC GGTTCCCAAC
6421 GATCAAGGCG AGTTACATGA TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC
6481 CTCCGATCGT TGTCAGAAGT AAGTTGGCCG CAGTGTTATC ACTCATGGTT ATGGCAGCAC
6541 TGCATAATTC TCTTACTGTC ATGCCATCCG TAAGATGCTT TTCTGTGACT GGTGAGTACT
6601 CAACCAAGTC ATTCTGAGAA TAGTGTATGC GGCGACCGAG TTGCTCTTGC CCGGCGTCAA
6661 TACGGGATAA TACCGCGCCA CATAGCAGAA CTTTAAAAGT GCTCATCATT GGAAAACGTT
6721 CTTCGGGGCG AAAACTCTCA AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA
6781 CTCGTGCACC CAACTGATCT TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA
6841 AAACAGGAAG GCAAAATGCC GCAAAAAAGG GAATAAGGGC GACACGGAAA TGTTGAATAC
6901 TCATACTCTT CCTTTTTCAA TATTATTGAA GCATTTATCA GGGTTATTGT CTCATGAGCG
6961 GATACATATT TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC ACATTTCCCC
7021 GAAAAGTGCC ACCTGGG
pCXWB-EBNA1(Plasmid #37624)
pCXWB-EBNA1(Plasmid #37624)
pCXWB-EBNA1(Plasmid #37624)
