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pBI series are binary vectors with pBI vector backbone and Gateway® cassette for cloning. pBI series includes various vectors for your experiments, gene expression with any promoter, overexpression, and RNA silencing. Vectors with GFP as reporter gene, and vectors with specific sequences to distinguish introduced gene from native one at RT-PCR, are also available.
【IN3-VEC1】pBI-GW-NOS promoter less type
pBI-GW-NOS is a Gateway® binary vector for making expression construct with any promoter and preparation of a complementation construct with genome fragment. This vector has been practically used in RIKEN Plant Science Center.
Features
・Promoter less Gateway® cassette is connected at upstream of NOS terminator.
・E.coli and Agrobacterium selection marker: NPTIII (kanamycin resistant)
・Plant selection marker: NPTII (kanamycin resistant)
Enzyme map(PDF) Sequence(Fasta) Sequence (PDF)
Reference (used as pBCR112 in the reference)
M Suzuki et al., J. Exp. Bot. (2009) 60 (7): 2055-2064.
J Tang et al.,Plant J. (2010)61(3):456-466.
【IN3-VEC2】pBI-OX-GW overexpression type
pBI-OX-GW is a Gateway® binary vector for making overexpression construct and used for gene function analysis and preparation of transgenic plants with overexpression of purpose gene. This vector has been practically used in RIKEN Plant Science Center.
Features
・High expression by double enhancers with CaMV35S promoter
・Gateway® cassette is available between 35SS promoter and NOS terminator.
・E.coli and Agrobacterium selection marker: NPTIII (kanamycin resistant)
・Plant selection marker: NPTII (kanamycin resistant)
Enzyme map(PDF) Sequence(Fasta) Sequence(PDF)
References (Used as pBCR79 in the references)
K Kobayashi et al., Plant Cell Physiol (2007) 48 (2):322-331.
K Nemoto et al., J Plant Phys (2009)166(7):729-738.
K Nemoto et al., FEBS Let(2009)583(2):487-492.
【IN3-VEC3】pBI-sense, anti sense-GW RNA silencing type
pBI-sense, anti sense-GW is a Gateway® binary vector for RNA silencing (RNAi). This vector is useful for the purpose of gene knock-down. The vector has been practically used in RIKEN Plant Science Center.
Features
・High expression of the Gateway® cassette inverted repeat which has an intron sequence at the center by the double enhancer CaMV35S promoter.
・Only one time LR reaction is necessary for preparing of RNAi construct of your purpose gene.
・E.coli and Agrobacterium selection marker: NPTIII (kanamycin resistant)
・Plant selection marker: NPTII (kanamycin resistant)
Enzyme map(PDF) Sequence(Fasta) Sequence (PDF)
Reference (Used as pBCR80 in the reference)
K Nemoto et al., FEBS Let. (2009)583(2):487-492.
【IN3-VEC4】pBI-GW-NOS (GFP selection) promoter less/GFP reporter type
pBI-GW-NOS (GFP selection) is a promoter less Gateway® binary vector which has GFP as a reporter, for making expression construct with any promoter and preparation of a complementation construct with genome fragment. This vector has been practically used in RIKEN Plant Science Center.
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Features
・Promoter less Gateway® cassette is connected at upstream of NOS terminator.
・E.coli and Agrobacterium selection marker: NPTIII (kanamycin resistant)
・Plant selection marker: NPTII (kanamycin resistant)
・Selection of transgenic plant by sGFP(S65T) fluorescence
Enzyme map(PDF) Sequence(Fasta) Sequence(PDF)
【IN3-VEC5】pBI-OX-GW (GFP selection) overexpression type/GFP reorter type
pBI-OX-GW(GFP selection) is a Gateway® binary vector harboring GFP as a reporter for making overexpression construct and used for gene function analysis and preparation of transgenic plants with overexpression of purpose gene. This vector has been practically used in RIKEN Plant Science Center.
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Features
・High expression by double enhancers with CaMV35S promoter
・Gateway® cassette is available between 35SS promoter and NOS terminator.
・Selection of transgenic plant by sGFP(S65T) fluorescence
・E.coli and Agrobacterium selection marker: NPTIII (kanamycin resistant)
・Plant selection marker: NPTII (kanamycin resistant)
Enzyme map(PDF) Sequence(Fasta) Sequence(PDF)
【IN3-VEC6】pBI-sense,anti sense-GW (GFP selection)
RNA silencing/GFP reporter type
pBI-sense, anti sense-GW (GFP selection) is a Gateway® binary vector harboring GFP as a reporter for RNA silencing (RNAi). This vector is useful for the purpose of gene knock-down. The vector has been practically used in RIKEN Plant Science Center.
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Features
・High expression of the Gateway® cassette inverted repeat which has an intron sequence at the center by the double enhancer CaMV35S promoter.
・Only one time LR reaction is necessary for preparing of RNAi construct of your purpose gene.
・Selection of transgenic plant by sGFP(S65T) fluorescence
・E.coli and Agrobacterium selection marker: NPTIII (kanamycin resistant)
・Plant selection marker: NPTII (kanamycin resistant)
Enzyme map(PDF) Sequence(Fasta) Sequence(PDF)
【IN3-VEC7】pBIDAVL-GWR1
Sence overexpression/with specific sequence for RT-PCR type
pBIDAVL-GWR1 is a Gateway® binary vector for making sense overexpression construct with a unique sequence for RT-PCR primer (DAVL) and used for gene function analysis and preparation of transgenic plants with overexpression of purpose gene. This vector has been practically used in RIKEN Plant Science Center.
Features
・High expression by double enhancers with CaMV35S promoter
・Gateway® cassette is available between 35SS promoter and NOS terminator.
・Easy check of accumulation of introduced gene by RT-PCR with a primer which can be set in the unique sequence (DAVL) locating before polyA signal and gene specific primer.
・Only one entry clone is required to make both sense and antisense constructs by using pBIDAVL-GWR2
・E.coli and Agrobacterium selection marker: NPTIII (kanamycin resistant)
・Plant selection marker: NPTII (kanamycin resistant)
Enzyme map(PDF) Sequence(Fasta) Sequence(PDF)
【IN3-VEC8】pBIDAVL-GWR2
Antisence overexpression/with specific sequence for RT-PCR type
pBIDAVL-GWR2 is a Gateway® binary vector for making antisense overexpression construct with a unique sequence for RT-PCR primer (DAVL) and used for gene function analysis and preparation of transgenic plants with overexpression of purpose gene. This vector has been practically used in RIKEN Plant Science Center.
Features
・High expression of antisense gene by CaMV35S promoter
・Gateway® cassette is available between 35SS promoter and NOS terminator.
・Easy check of accumulation of introduced gene by RT-PCR with a primer which can be set in the unique sequence (DAVL) locating before polyA signal and gene specific primer.
・Only one entry clone is required to make both sense and antisense constructs by using pBIDAVL-GWR1
・E.coli and Agrobacterium selection marker: NPTIII (kanamycin resistant)
・Plant selection marker: NPTII (kanamycin resistant)
Size and buffer
Size : 10µg
Buffer : 10mM Tris-HCl(pH7.4) ， 1mM EDTA
How to use
1. Prepare entry clone which harboring your target gene. ※1
Some methods are available to make entry clones though, we recommend to use BP reaction※2 with PCR products and donor vectors. ※3
(Selection of E. coli is dependent on antibiotic resistance of donor vector. Competent cells of DH5α※4 with high efficiency over 1010 are recommended.)
2. Prepare expression clone by LR ※4 reaction with entry clone which was prepared in the step 1 and destination vector.
(Kanamycin should be used for selection. Competent cells of DH5α※5 with high efficiency over 10^10 are recommended.)
3. Transform the expression clone which was prepared at step 2. into Agrobacterium and infect them to plants.
(Transgenic plants must be selected by antibiotics which is designated for each vector. )
※1 See invitrogen HP for in detail.
※2 Invitrogen Gateway BP Clonase Enzyme Mix
※3 Invitrogen pDONR Gateway/Zeo vector(Zeocin resistance)
※4 Invitrogen Gateway LR Clonase Enzyme Mix
※5 Invitrogen Library Efficiency DH5α Competent cells
Fusion construct
・For fusion construct of your purpose gene and the fluorescent protein gene, please adjust codon frame of both genes.
License and guarantee
・Gateway® system is Trade mark of Invitrogen Inc. A license agreement is required for purchase and use of this vector.
・The vectors have been functionally estimated in plant transformation test in RIKEN and transformed into E coli., amplified and purified in Inplanta Innovations Inc.
・Other purpose of transformation of plants are not warranted, for example, using of fragments cut out from vectors.
・Listed products are intended for laboratory research use only.
All rights reserved - Inplanta Innovations Inc.