Gateway Binary Vectors with the Bialaphos Resistance Gene,<i>bar</i>, as a Selection Marker for Plant Transformation

Nakamura, Shinya; Mano, Shoji; Tanaka, Yuji; Ohnishi, Masato; Nakamori, Chihiro; Araki, Masami; Niwa, Takeo; Nishimura, Mikio; Kaminaka, Hironori; Nakagawa, Tsuyoshi; Satō, Yutaka; Ishiguro, Sumie

doi:10.1271/bbb.100184

Cited by 205 publications

(183 citation statements)

References 29 publications

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“…The plasmid constructs were produced using Gateway Technology (Invitrogen) with the destination vectors pGWB405 (Nakagawa et al, 2007), pGWB560 (AB543177; Nakagawa et al, 2007;Nakamura et al, 2010), pBGWF7 (Karimi et al, 2002;Plant System Biology), and FAST-R7 Plant System Biology). The CLO3 complementary DNA (cDNA) fragment from nucleotide 1, corresponding to A of the start codon ATG, to nucleotide 708 was amplified by PCR using the primer set 59-CACCATGGCAG-GAGAGGCAGAGGCTT-39 and 59-GTCTTGTTTGCGAGAATTGGCCC-39.…”

Section: Plasmid Dna Constructsmentioning

confidence: 99%

Leaf Oil Body Functions as a Subcellular Factory for the Production of a Phytoalexin in Arabidopsis

et al. 2013

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Section: Plasmid Dna Constructsmentioning

confidence: 99%

Leaf Oil Body Functions as a Subcellular Factory for the Production of a Phytoalexin in Arabidopsis

et al. 2013

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“…Pnos, nopaline synthase promoter; Tnos, nopaline synthase terminator; P 35S , 35S promoter; NPTII, neomycin phosphotransferase II; HPT, hygromycin phophotransferase; bar, bialaphos resistance gene; GPT, UDP-Nacetylglucosamine: dolichol phosphate N-acetylglucosamine-1-P transferase (Koizumi & Iwata, 2008;Koizumi et al, 1999) gene. Km r , kanamycin-resistance; Hyg r , hygromycinresistance; Spc r , spectinomycin-resistance; BASTA ®r , BASTA ® -resistance; Tunicamycin r , tunicamycin-resistance At present, four kinds of ImpGWB, the Km r subseries (pGWB4xx) (Nakagawa et al, 2007b), Hyg r subseries (pGWB5xx) (Nakagawa et al, 2007b), BASTA ® -resistance subseries (pGWB6xx) (Nakamura et al, 2010) and tunicamycin-resistance subseries (pGWB7xx) , are available, and they are useful for introducing multiple transgenes into plants by repetitive transformation. Each subseries is composed of 46 vectors as summarized in the Complete List of ImpGWB (http:/ / shimane-u.org/ nakagawa/ gbv.htm).…”

Section: The Pgwb Series (Pgwbxx and Pgwb2xx) Based On The Pbi Plasmidmentioning

confidence: 99%

Gateway Vectors for Plant Genetic Engineering: Overview of Plant Vectors, Application for Bimolecular Fluorescence Complementation (BiFC) and Multigene Construction

Tanaka¹,

Kimura²,

Hikino³

et al. 2012

Genetic Engineering - Basics, New Applications and Responsibilities

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“…All entry clones were recombined into the Gateway destination vector pGWB602V (Nakamura et al, 2010) via LR reactions using Gateway LR Clonase II enzyme mix (Invitrogen), to generate the corresponding binary vectors expressing 35S:MYB. Binary vectors were verified by restriction-enzyme digestion.…”

Section: Generation Of 35s:myb and 35s:mmyb Constructsmentioning

confidence: 99%

Target RNA Secondary Structure Is a Major Determinant of miR159 Efficacy

et al. 2017

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In plants, microRNA (miRNA)-target complementarity has long been considered the predominant factor determining the silencing outcome of the miRNA-target interaction, although the efficacy of such interactions have rarely been appraised in plants. Here, we perform in planta silencing efficacy assays on seven Arabidopsis MYB genes, all of which contain conserved miR159-binding sites of analogous complementarity. These genes were found to be differentially silenced by miR159; MYB81, MYB97, MYB101, MYB104, and DUO1 were all poorly silenced, whereas MYB33 and MYB65 were strongly silenced. Curiously, this is consistent with previous genetic analysis defining MYB33 and MYB65 as the major functional targets of miR159. Neither the free energy of miR159-target complementarity, nor miRNA binding site accessibility, as determined by flanking region AU content, could fully explain the discrepancy of miR159 silencing efficacy. Instead, we found that MYB33 and MYB65 were both predicted to contain a distinctive RNA secondary structure abutting the miR159 binding site. The structure is composed of two stem-loops (SLs) that are predicted to form in MYB33/65 homologs of species as evolutionary distant as gymnosperms. Functional analysis found that the RNA structure in MYB33 correlated with strong silencing efficacy; introducing mutations to disrupt either SL attenuated miR159 efficacy, while introducing complementary mutations to restore the SLs, but not the sequence, restored strong miR159-mediated silencing. Therefore, it appears that this RNA secondary structure demarcates MYB33/65 as sensitive targets of miR159, which underpins the narrow functional specificity of Arabidopsis miR159.

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Gateway Binary Vectors with the Bialaphos Resistance Gene,bar, as a Selection Marker for Plant Transformation

Cited by 205 publications

References 29 publications

Leaf Oil Body Functions as a Subcellular Factory for the Production of a Phytoalexin in Arabidopsis

Leaf Oil Body Functions as a Subcellular Factory for the Production of a Phytoalexin in Arabidopsis

Gateway Vectors for Plant Genetic Engineering: Overview of Plant Vectors, Application for Bimolecular Fluorescence Complementation (BiFC) and Multigene Construction

Target RNA Secondary Structure Is a Major Determinant of miR159 Efficacy

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