The 35S promoter used in a selectable marker gene of a plant transformation vector affects the expression of the transgene

Yoo, So Yeon; Bomblies, Kirsten; Yoo, Seung Kwan; Yang, Jung Won; Choi, Myung Suk; Lee, Jong Seob; Weigel, Detlef; Ahn, Ji Hoon

doi:10.1007/s00425-004-1466-4

Cited by 147 publications

(106 citation statements)

References 30 publications

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“…This fragment was then cloned into the BglII/BamHI sites of pURTkan, a derivative of pJHA212 (Yoo et al, 2005), containing the Ubiquitin 10 promoter and the mRFP coding sequence. The resulting binary plasmid was introduced into Agrobacterium tumefaciens strain GV3101:pMP90 and selected on 5 mg/mL rifampicin, 10 mg/mL gentamycin, and 100 mg/mL spectinomycin.…”

Section: Plasmid Constructs and Plant Transformationmentioning

confidence: 99%

Multivesicular Bodies Mature from the Trans-Golgi Network/Early Endosome in Arabidopsis

et al. 2011

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The plant trans-Golgi network/early endosome (TGN/EE) is a major hub for secretory and endocytic trafficking with complex molecular mechanisms controlling sorting and transport of cargo. Vacuolar transport from the TGN/EE to multivesicular bodies/late endosomes (MVBs/LEs) is assumed to occur via clathrin-coated vesicles, although direct proof for their participation is missing. Here, we present evidence that post-TGN transport toward lytic vacuoles occurs independently of clathrin and that MVBs/LEs are derived from the TGN/EE through maturation. We show that the V-ATPase inhibitor concanamycin A significantly reduces the number of MVBs and causes TGN and MVB markers to colocalize in Arabidopsis thaliana roots. Ultrastructural analysis reveals the formation of MVBs from the TGN/EE and their fusion with the vacuole. The localization of the ESCRT components VPS28, VPS22, and VPS2 at the TGN/EE and MVBs/LEs indicates that the formation of intraluminal vesicles starts already at the TGN/EE. Accordingly, a dominant-negative mutant of VPS2 causes TGN and MVB markers to colocalize and blocks vacuolar transport. RNA interference-mediated knockdown of the annexin ANNAT3 also yields the same phenotype. Together, these data indicate that MVBs originate from the TGN/EE in a process that requires the action of ESCRT for the formation of intraluminal vesicles and annexins for the final step of releasing MVBs as a transport carrier to the vacuole.

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Section: Plasmid Constructs and Plant Transformationmentioning

confidence: 99%

Multivesicular Bodies Mature from the Trans-Golgi Network/Early Endosome in Arabidopsis

et al. 2011

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“…Most likely, the discrepancy is caused by a double-enhanced cauliflower mosaic virus 35S promoter located adjacent to the EXO70A1 promoter in the pCAMBIA1391Z vector used by Samuel et al (2009). It has been reported previously that the active 35S promoter in some of the pCAMBIA vectors including pCAMBIA1391Z causes significant interference to the expression patterns of the promoters analyzed (Yoo et al, 2005; http://www. patentlens.net/daisy/cambia/home.html).…”

Section: Genes In Arabidopsismentioning

confidence: 99%

Expression and Functional Analyses ofEXO70Genes in Arabidopsis Implicate Their Roles in Regulating Cell Type-Specific Exocytosis

Ren

et al. 2010

Plant Physiology

107

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During exocytosis, Golgi-derived vesicles are tethered to the target plasma membrane by a conserved octameric complex called the exocyst. In contrast to a single gene in yeast and most animals, plants have greatly increased number of EXO70 genes in their genomes, with functions very much unknown. Reverse transcription-polymerase chain reactions were performed on all 23 EXO70 genes in Arabidopsis (Arabidopsis thaliana) to examine their expression at the organ level. Cell-level expression analyses were performed using transgenic plants carrying β-glucuronidase reporter constructs, showing that EXO70 genes are primarily expressed in potential exocytosis-active cells such as tip-growing and elongating cells, developing xylem elements, and guard cells, whereas no expression was observed in cells of mature organs such as well-developed leaves, stems, sepals, and petals. Six EXO70 genes are expressed in distinct but partially overlapping stages during microspore development and pollen germination. A mutation in one of these genes, EXO70C1 (At5g13150), led to retarded pollen tube growth and compromised male transmission. This study implies that multiplications of EXO70 genes may allow plants to acquire cell type- and/or cargo-specific regulatory machinery for exocytosis.

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“…Details are provided in the Supplemental Methods online. These constructs were subcloned into the pJHA212K binary vector (a gift from Ji Hoon Ahn) (Yoo et al, 2005) and introduced into Agrobacterium tumefaciens C58C1 or ASE. Arabidopsis plants (Col-0 wild type, ttg2-1, and ttg2-3) were transformed by the floral dipping method and screened on 0.8% agar plates containing half-concentration Murashige and Skoog medium and 50 mg/L kanamycin sulfate.…”

Section: Constructs and Plant Transformationmentioning

confidence: 99%

Arabidopsis TRANSPARENT TESTA GLABRA2Is Directly Regulated by R2R3 MYB Transcription Factors and Is Involved in Regulation ofGLABRA2Transcription in Epidermal Differentiation

et al. 2007

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Arabidopsis thaliana TRANSPARENT TESTA GLABRA2 (TTG2) encodes a WRKY transcription factor and is expressed in young leaves, trichomes, seed coats, and root hairless cells. An examination of several trichome and root hair mutants indicates that MYB and bHLH genes regulate TTG2 expression. Two MYB binding sites in the TTG2 59 regulatory region act as cis regulatory elements and as direct targets of R2R3 MYB transcription factors such as WEREWOLF, GLABRA1, and TRANSPARENT TESTA2. Mutations in TTG2 cause phenotypic defects in trichome development and seed color pigmentation. Transgenic plants expressing a chimeric repressor version of the TTG2 protein (TTG2:SRDX) showed defects in trichome formation, anthocyanin accumulation, seed color pigmentation, and differentiation of root hairless cells. GLABRA2 (GL2) expression was markedly reduced in roots of ProTTG2:TTG2:SRDX transgenic plants, suggesting that TTG2 is involved in the regulation of GL2 expression, although GL2 expression in the ttg2 mutant was similar to that in the wild type. Our analysis suggests a new step in a regulatory cascade of epidermal differentiation, in which complexes containing R2R3 MYB and bHLH transcription factors regulate the expression of TTG2, which then regulates GL2 expression with complexes containing R2R3 MYB and bHLH in the differentiation of trichomes and root hairless cells.

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The 35S promoter used in a selectable marker gene of a plant transformation vector affects the expression of the transgene

Cited by 147 publications

References 30 publications

Multivesicular Bodies Mature from the Trans-Golgi Network/Early Endosome in Arabidopsis

Multivesicular Bodies Mature from the Trans-Golgi Network/Early Endosome in Arabidopsis

Expression and Functional Analyses ofEXO70Genes in Arabidopsis Implicate Their Roles in Regulating Cell Type-Specific Exocytosis

Arabidopsis TRANSPARENT TESTA GLABRA2Is Directly Regulated by R2R3 MYB Transcription Factors and Is Involved in Regulation ofGLABRA2Transcription in Epidermal Differentiation

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