pRI909载体简介:
Binary Vectors for Agrobacterium-mediated Plant Transformation: pRI909 and pRI910
Ri Plasmid pRI909 and pRI910 are binary vectors intended for Agrobacterium-mediated plant
transformation and contain a region of T-DNA for Arabidopsis, tomato, tobacco and rice
transformation. The origin of the T-DNA is the vir region-lacking Rhizobium (Agrobacterium)
rhizogenes Ri plasmid. Ri Plasmid pRI909 and pRI910 are also shuttle vectors and replicate
autonomously in E. coli and Rhizobium (Agrobacterium). In E. coli, these vectors are capable
of high copy number because they contain the ColE1 replication origin. The ColE1 ori, as well
as the mutant-type Ri plasmid replication of origin (Ri-ori), are each stably maintained in
Rhizobium (Agrobacterium). These vectors also contain the kanamycin resistance gene NPTIII
for selection in E. coli and Rhizobium (Agrobacterium) and a mutant-type kanamycin resistance
gene NPTII for selection in plants.
The Ri Plasmid pRI909 and pRI910 vectors are capable of plant transformation in combination
with Rhizobium (Agrobacterium) through a binary vector system. Stable plant chromosome target
gene integration is possible with the Ri Plasmid pRI909 and pRI910 because their cloning
sites are located in close proximity to the T-DNA Right Border(RB) and not the NPT II plant
selection marker. As a result, the target gene is not deleted.
pRI909载体特征:
Easy and stable plant chromosome target gene integration
Improved agrobacterium-mediated plant transformation
Versatile: Applicable to various plants such as Arabidopsis, tomato, tobacco, and rice
Applications
Rhizobium (Agrobacterium)-mediated plant transformation
Arabidopsis, tomato, tobacco, rice transformation
Purity
>70% double-stranded covalently closed circular form I (RFI) DNA.
Cloning sites verified by dideoxy DNA sequencing.
Restriction sites verified by restriction enzyme cleavage.
参考文献:
1. Nishiguchi, R. et al. (1987) Mol. Gen. Genet. 206:1-8.
2. Ooms, G. et al. (1982) Plasmid 7:15-29.
3. Ohba, T. et al. (1995) The Plant J. 7:157-164.
TGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTC
CATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGA
AACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCT
CCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTG
GCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAG
CTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTAT
CGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAAC
AGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAAC
TACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTC
GGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTT
TTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATC
TTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATG
AGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCA
ATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCA
CCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCGATCCTACAAGG
TAGAATCCGCCTGAGTCGCAAGGGTGACTTCGCCTATATTGGACGACGGCGCGCAGAGGG
CGACCTCTTTTTGGGTTACGATTGTAGGATTATCACTAAAACAATACATGAACATATTCA
AATGGCAATCTCTCTAAGGCATTGGAAATAAATACAAATAACAGTTGGGTGGAGTTTTTC
GACCTGAGGGCGTTAACCTTCTGTTAACCTAAAAGCTCTTGCCCAAACAGCAGAATCGGC
GCTAATTGCCAGCGGCGGAACTTTTCCAGTTTCGCGAAAAATATCGCCACTGGCAAGGAA
TGGGTTTGAGATGGCGAAGTCTGTCCTAAAAGCAGCGCCTGTAGTTGTAGGGTTGACGGC
CTTGATGGAGCGTCATGCCGATGCCCTCTCGAGCCAACTTCAAGCACATCATCTTAAGGT
TTTCCCGCCGCATTCCGAGAAGGGTATTCGAACATTCGGGCCATCGGAGGCGTCCAAGCT
GCTCGGCGTTGGCGAGTCATATTTACGGCAGACCGCGTCTGAGATGCCAGAGTTGAATGT
TAGCATGAGCCCAGGTGGCAGGCGAATGTTCTCAATTGAAGATATCCATGTGATTCGGAA
GTATATGGATCAGGTCGGCCGCGGGAACCGGCGCTACCTGCCACATCGTCGAGGCGGCGA
GCAGCTTCAGGTTATCTCTGTGATGAATTTCAAAGGTGGGTCGGGTAAGACCACCACCGC
CGCGCATCTGGCGCAGTACCTCGCTATGCGCGGATATCGAGTCTTGGCCATTGATCTCGA
TCCTCAAGCGAGCCTTTCTGCACTCTTTGGGAGCCAACCGGAGACGGACGTTGGCCCGAA
CGAAACGCTCTACGGCGCTATAAGGTATGATGATGAGCAGGTGGCAATCGAACGAGTCGT
CCGAGGGACTTACATTCCCGACCTCCACCTGATTCCTGGTAACCTTGAGCTGATGGAGTT
TGAACACGATACGCCACGCGCGCTGATGAACCGCAAAGAGGGCGACACGCTCTTTTATGG
TCGCATCAGCCAAGTAATTGAAGATATCGCGGATAACTATGACGTCGTGGTCATCGACTG
CCCTCCCCAGCTTGGGTATCTCACGCTATCCGCATTGACTGCGGCGACGTCCATTCTTGT
CACGGTCCATCCGCAGATGCTGGATGTGATGTCGATGAACCAGTTTCTGGCAATGACATC
GAACCTTTTGCGTGAAATCGAGAATGCTGGCGCCAAGTTCAAGTTTAATTGGATGCGCTA
TCTGATAACCCGTTTCGAACCGAGCGACGGACCACAGAACCAAATGGTAGGTTATCTGCG
GTCGATTTTTGGCGAAAATGTCCTCAATTTTCCGATGCTTAAAACCACCGCGGTTTCGGA
CGCTGGCCTGACAAACCAGACTCTATTCGAAGTGGAGCGTGGCCTGTTCACGCGCTCGAC
CTATGATCGAGCCTTGGAGGCGATGAACGCCGTCAACGACGAGATCGAAACACTGATCAA
AAAAGCATGGGGTAGGCCCACATGAGCCGGAAGCACATCCTTGGCGTCTCAACTGACGCC
CCTGAGACGTCGCCCGCCGACAATAGGACGGCAAAGAACCGCTCCATGCCGCTCCTCGGC
GTAACAAGGAAGGAGCGCGATCCGGCAACGAAGCTCACAGCGAACATTGGTAACGCACTG
CGAGAGCAAAACGATCGTCTTAGCCGTGCCGAAGAGATCGAGCGGCGTCTCGCTGAAGGT
CAGGCAGTGATAGAGTTGGATGCCTCGTCAATAGAACCGTCTTTCGTGCAGGATCGTATG
CGAGGGGACATTGACGGGCTCCTTACTTCGATCCGGGAACAAGGACAGCAAGTCCCAATC
CTTGTGCGACCGCATCCGAGCCAGCCGGGCCGATATCAGGTTGCCTTCGGCCACCGCCGG
CTACGCGCCGTTTCAGAACTCGGACTTCCGGTCAGGGCGGTCGTTCGCGAACTGACGGAC
GAGCAAGTGGTCGTAGCACAGGGTCAGGAAAACAATGTGCGCGAAGATCTTACCTTCATC
GAAAAGGCGCGCTTCGCACATCGCCTGAACAGGCAGTTTTCTCGAGAGATTGTCATCGCC
GCGATGTCGATCGACAAGAGCAATTTGTCCAAGATGCTTCTGCTCGTTGACGCCCTCCCC
TCTGAACTGACCGATGCTATTGGTGCCGCTCCTGGTGTTGGACGGCCGAGTTGGCAACAA
CTTGCCGAGCTGATTGAGAAAGTTTCTTCACCGGCCGACGTGGCTAAATATGCTATGTCG
GAGGAAGTTCAAGCGCTGCCATCGGCAGAACGATTCAAGGCGGTGATCGCTAGTCTGAAG
CCCAGTCGGGTTGCGCGTGGACTTCCCGAGGTCATGGCCACCCCAGACGGCACCAGAATT
GCACAGGTGACGCAGAGCAAGGCCAAACTGGAAATCACGATTGACAGGAAGGCGACGCCC
GATTTTGCGACCTTCGTGCTCGATCATGTGCCAGCGCTGTATCAAGCGTACCACGCTGAG
AACCAACGGAAACGGGGAGAGTAAACCGCAAAAGAAAAGAGCCCCCTCAACGTCGCCGTC
GCGGAAGCCCTTCTGTCTCTCTAGCGCGAACAGAATCGCATTTCCTCGAATCCTCGTCAA
GAGTTTTTAGCGCCGTTTTGGTGAGCTGATTTCCTTTGCCTGCTGAAAGGTGAAAGATGA
TGCAGACAGGAAGTGTAACGACGCCATTCGGGCGGCGGCCAATGACGCTTGCGCTTGTGC
GGCGCCAGACGGCGCTGGCCGATATCAAACAAGGCAAGACAGCGGACAAGTGGAAGGTCT
TTAGGGACGCGTCCGCGGCCATGGAACTACTTGGAATCCAGTCCAACAGTCTTGCCGTCC
TTGATGCGCTATTGAGCTTTCACCCGGAAACGGAGTTGCGTCAGGAGGCACAGCTGATCG
TCTTCCCGTCGAATGCTCAGCTTGCCCTTCGGGCGCATGGGATGGCTGGCGCGACTTTGC
GTAGGCACATCGCCATGCTCGTGGAGTCAGGCTTGATCGTCCGGAAGGATAGCGCCAACG
GAAAGCGTTACGCTCGTAAGGATGGCGCTGGTCAGATCGAGCGCGCGTTTGGCTTCGATT
TGTCTCCGCTTCTCGCGCGGTCCGAAGAGCTAGCGATGATGGCACAGCAGGTGATGGCCG
ATCGAGCAGCATTCAGGATGGCCAAAGAAAGTCTGACGATTTGCCGACGGGACGTTCGGA
AGCTAATTACGGCAGCTATGGAAGAGGGAGCGGAGGGCGACTGGCAAGCTGTCGAGGAAG
TCTATGTGGAACTTGTGGGTAGAATCCCACGCGCCCCGACGCTTGCTGATGTAGAGTCAA
TTCTCGAAGAGATGTGGATGCTCCAGGAAGAGATAATCAACCGGTTGGAAATTAGAGACA
ATTCAGAAAATAATAGCACCAATGCTGCCCAGAGCGAGCAGCACATACAGAACTCAAAAC
CCGAATCCGTTAATGAACTTGAACCTCGCTCTGAAAAGGAGCAGGGCGCTAAGCCGAGTG
AAATAGACCGGGCAAGGAGCGAGCCGATAAAAGCGTTCCCCCTCGGGATGATCCTGAAAG
CGTGCCCGACCATTGGCAATTATGGGCCGAGCGGTGCGGTTGCTAGCTGGCGTGACCTCA
TGTCGGCTGCGGTGGTGGTTCGGTCTATGCTGGGGGTCAGCCCGTCGGCTTACCAAGACG
CGTGTGAGGCAATGGGACCGGAGAATGCGGCAGCAGCGATGGCGTGCATTTTGGAGCGAG
CGAACTTCATCAATTCGCCTGGGGGCTATCTCCGAGATCTGACACGGCGGAGCGAACTCG
GGAAGTTTTCACTTGGCCCGATGATAATGGCGCTCTTGAAGGCTAGCGGGCAGGGGACGT
TGCGGTTTGGCTAGAATTAGCGAGTATGGAGCAGGATGGTCTGTGGTCAGCTGACCACAG
ACCTAATAGGTTGAAAACATGAGCGTTTTTTGGATGATCGACAGACCATCCGATTCCCGG
AGTACCAAGCGTGCTCTGATGGGAGCGATAACATTACTCAACAAGCACGAAGGCCCCATG
CCGATCGTTGATCGTGAAGGAGAGCCTGCTCTACATGCGGCGGTATTTTGCCGGCCGAGG
CATGTAGTCGCGGAGCACTGCCTATTTACTGCCCTAGGCACAAACGTTGACTCTTGGATC
GAGCTGGCAGACAAAGCAATAACCCACACAGAGGACGATTAATGGCTGACGAAGAGATCC
AGAATCCGCCGGACGGTACTGCTGCTGCCGAAGTTGAGCCGGCTGCTCCTAGAGGTAGAA
GAGCAAAGAAAGCACCAGCCGAAACAGCCCGCACGGGATCGTTCAAATCCGTGAAGCCGA
AAACCCGCGGCCTCAGCAACCGAGAAAAACTGGAGAAGATCGGTCAAATCGAAGCTCAGG
TCGCTGGCGGCGCAACCTTGAAGGACGCCGTTAAGATCGTGGGTATTTCCGTTCAGACCT
ATTATCAATGGAAGAGAGCTGCGGTTCAACCTGTCTCACAGAATCCGGCCGTGTCTGTTT
CAGTTGACGATGAACTCGGCGAGTTCATCCAACTCGAGGAGGAAAATCGGCGGCTCAGAA
AGCTAGCTTGTGGCAGGATATATTGTGGTGTAAACAAATTGACGCTTAGACAACTTAATA
ACACATTGCGGACGTTTTTAATGTACTGGGGTGGTTTTTCTTTTCACCAGTGAGACGGGC
AACAGCTGATTGCCCTTCACCGCCTGGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTG
GTTTGCCCCAGCAGGCGAAAATCCTGTTTGATGGTGGTTCCGAAATCGGCAAAATCCCTT
ATAAATCAAAAGAATAGCCCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTC
CACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATG
GCCCAAATTCCCGATCTAGTAACATAGATGACACCGCGCGCGATAATTTATCCTAGTTTG
CGCGCTATATTTTGTTTTCTATCGCGTATTAAATGTATAATTGCGGGACTCTAATCATAA
AAACCCATCTCATAAATAACGTCATGCATTACATGTTAATTATTACATGCTTAACGTAAT
TCAACAGAAATTATATGATAATCATCGCAAGACCGGCAACAGGATTCAATCTTAAGAAAC
TTTATTGCCAAATGTTTGAACGATCGGGGCCCGATCATATTGTCGCTCAGGATCGTGGCG
TTGTGCTTGTCGGCCGTTGCTGTCGTAATGATATCGGCACCTTCGACCGCCTGTTCCGCA
GAGATCCCGTGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCG
GCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAA
TCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTC
AGAAGAACTCGTCAAGAAGGCGATAGAAGGCGATGCGCTGCGAATCGGGAGCGGCGATAC
CGTAAAGCACGAGGAAGCGGTCAGCCCATTCGCCGCCAAGCTCTTCAGCAATATCACGGG
TAGCCAACGCTATGTCCTGATAGCGGTCCGCCACACCCAGCCGGCCACAGTCGATGAATC
CAGAAAAGCGGCCATTTTCCACCATGATATTCGGCAAGCAGGCATCGCCATGGGTCACGA
CGAGATCATCGCCGTCGGGCATACGCGCCTTGAGCCTGGCGAACAGTTCGGCTGGCGCGA
GCCCCTGATGCTCTTCGTCCAGATCATCCTGATCGACAAGACCGGCTTCCATCCGAGTAC
GTGCTCGCTCGATGCGATGTTTCGCTTGGTGGTCGAATGGGCAGGTAGCCGGATCAAGCG
TATGCAGCCGCCGCATTGCATCAGCCATGATGGATACTTTCTCGGCAGGAGCAAGGTGAG
ATGACAGGAGATCCTGCCCCGGCACTTCGCCCAATAGCAGCCAGTCCCTTCCCGCTTCAG
TGACAACGTCGAGCACAGCTGCGCAAGGAACGCCCGTCGTGGCCAGCCACGATAGCCGCG
CTGCCTCGTCCTGTAGTTCATTCAGGGCACCGGACAGGTCGGTCTTGACAAAAAGAACCG
GGCGCCCCTGCGCTGACAGCCGGAACACGGCGGCATCAGAGCAGCCGATTGTCTGTTGTG
CCCAGTCATAGCCGAATAGCCTCTCCACCCAAGCGGCCGGAGAACCTGCGTGCAATCCAT
CTTGTTCAATCATGCGAAACGATCCAGATCCGGTGCAGATTATTTGGATTGAGAGTGAAT
ATGAGACTCTAATTGGATACCGAGGGGAATTTATGGAACGTCAGTGGAGCATTTTTGACA
AGAAATATTTGCTAGCTGATAGTGACCTTAGGCGACTTTTGAACGCGCAATAATGGTTTC
TGACGTATGTGCTTAGCTCATTAAACTCCAGAAACCCGCGGCTGAGTGGCTCCTTCAACG
TTGCGGTTCTGTCAGTTCCAAACGTAAAACGGCTTGTCCCGCGTCATCGGCGGGGGTCAT
AACGTGACTCCCTTAATTCTCCGCTCATGATCCTGGCGAAAGGGGGATGTGCTGCAAGGC
GATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCAGTG
CCAAGCTTGCATGCCTGCAGGTCGACTCTAGAGGATCCCCGGGTACCGAGCTCGAATTCG
TAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAAC
ATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACA
TTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGGATCAG
ATTGTCGTTTCCCGCCTTCAGTTTAAACTATCAGTGTTTGACAGGATATATTGGCGGGTA
AACCTAAGAGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAAGGGCGTGAAAAGGT
TTATCCGTTCGTCCATTTGTATGTGACTAAAACAATTCATCCAGTAAAATATAATATTTT
ATTTTCTCCCAATCAGGCTTGATCCCCAGTAAGTCAAAAAATAGCTCGACATACTGTTCT
TCCCCGATATCCTCCCTGATCGACCGGACGCAGAAGGCAATGTCATACCACTTGTCCGCC
CTGCCGCTTCTCCCAAGATCAATAAAGCCACTTACTTTGCCATCTTTCACAAAGATGTTG
CTGTCTCCCAGGTCGCCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTTTAAA
AAATCATACAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGTTTTCGCAATCC
ACATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCTAAGCGGCTGTCTAAGCTA
TTCGTATAGGGACAATCCGATATGTCGATGGAGTGAAAGAGCCTGATGCACTCCGCATAC
AGCTCGATAATCTTTTCAGGGCTTTGTTCATCTTCATACTCTTCCGAGCAAAGGACGCCA
TCGGCCTCACTCATGAGCAGATTGCTCCAGCCATCATGCCGTTCAAAGTGCAGGACCTTT
GGAACAGGCAGCTTTCCTTCCAGCCATAGCATCATGTCCTTTTCCCGTTCCACATCATAG
GTGGTCCCTTTATACCGGCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCCCACC
AGCTTATATACCTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGGTATTTTTCG
ATCAGTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATTATTTCCTTCCTCTT
TTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAACAAGACGAACTCCAATTCAC
TGTTCCTTGCATTCTAAAACCTTAAATACCAGAAAACAGCTTTTTCAAAGTTGTTTTCAA
AGTTGGCGTATAACATAGTATCGACGGAGCCGATTTTGAAACCACAATTATGGGTGATGC
TGCCAACTTACTGATTTAGTGTATGATGGTGTTTTTGAGGTGCTCCAGTGGCTTCTGTTT
CTATCAGCTGTCCCTCCTGTTCAGCTACTGACGGGGTGGTGCGTAACGGCAAAAGCACCG
CCGGACATCAGCGCTATCTCTGCTCTCACTGCCGTAAAACATGGCAAC