Multiple cis regulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants.

Fang, Rongxiang; Nagy, Ferenc; Sivasubramaniam, Shanthi; Chua, Nam‐Hai

doi:10.1105/tpc.1.1.141

Cited by 255 publications

(34 citation statements)

References 36 publications

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“…However, the repressive element was unable to suppress the activity of 35S enhancer in vegetative tissues [10,43]. In our study, the vegetative activity of the longer 351-bp 35S enhancer fragment (-396 to -46) that is responsible for the majority of the 35S promoter strength [33] could be suppressed by the repressive element of NtAGI-1, showing a potential of the repressive element in the design of synthetic reproductive promoters.…”

Section: Plos Onecontrasting

confidence: 52%

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A 100 bp GAGA motif-containing sequence in AGAMOUS second intron is able to suppress the activity of CaMV35S enhancer in vegetative tissues

Liu¹,

Zou

Wang³

et al. 2020

PLoS ONE

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Flower-specific promoters enable genetic manipulation of floral organs to improve crop yield and quality without affecting vegetative growth. However, the identification of strong tissuespecific promoters is a challenge. In addition, information on cis elements that is able to repress gene expression in vegetative tissues remains limited. Here, we report that fusing a 35S enhancer to the stamen-and carpel-specific NtAGIP1 promoter derived from the tobacco AGAMOUS second intron (AGI) can significantly increase the promoter activity. Interestingly, although the activity of the new promoter extends to sepals and pedicles, it does not cross the boundary of the reproductive organs. Serial deletion of the AGI and chromatin immunoprecipitation (ChIP) assay reveal a 100-bp fragment that contains a conserved GAGA factor binding motif contributes to the flower specificity by mediating histone H3 lysine 27 trimethylation (H3K27me3) modification of the promoter. Furthermore, this fragment shows significant suppressive effect on the activity of the 35S enhancer in vegetative tissues, consequently, resulting in a significant increase of the activity of 35S enhancer: AGI chimeric promoter without sacrifice of its specificity in inflorescence. A 100 bp GAGA motif-containing sequence in AGAMOUS second intron is able to suppress the activity of CaMV35S enhancer in vegetative tissues. PLoS ONE 15(3): e0230203. https://doi.org/ 10.The plasmid pBI121 [29] that contain CaMV35S::GUS was used as backbone to construct vectors used for plant transformation in this study. The second intron of NtAG-1 gene was fused with a 45 bp minimal 35S promoter to create a functional NtAGIP1 promoter [28]. NtAGIP1:: PLOS ONEA GAGA-containing sequence in AGAMOUS intron suppress the activity of CaMV35S enhancer in vegetative tissues PLOS ONE | https://doi.org/10.A GAGA-containing sequence in AGAMOUS intron suppress the activity of CaMV35S enhancer in vegetative tissues PLOS ONE | https://doi.org/10. Formal analysis: Ruochen Liu.Funding acquisition: Yan Pei.Investigation: Ruochen Liu, Xiuping Zou, You Wang, Qin Long. Fig 8. ChIP assay for H3K27me3 levels in NtAGI-1 intron and chimeric promoters. (A) ChIP assay for H3K27me3 levels at the NtAGI-1 intron in leaves of wild-type tobacco. (B) ChIP assay for H3K27me3 levels in leaves of the -2835 and -2735 lines. The genomic fragment from immunoprecipitation for qRT-PCR detection is depicted as black horizontal line. Enrichment was represented as percentage of Input (% Input). Error bars represent the standard deviation of three biological replicates.A GAGA-containing sequence in AGAMOUS intron suppress the activity of CaMV35S enhancer in vegetative tissues PLOS ONE | https://doi.org/10.

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Section: Plos Onecontrasting

confidence: 52%

“…To increase the promoter activity, we added 35S enhancer fragment (-396 to -46) [32,33] to the 5 'end of NtAGIP1, and generated 35SNtAGIP1 promoter ( Fig 1A). Then, NtAGIP1 and 35SNtAGIP1 were fused to the GUS coding region to create NtAGIP1::GUS and 35SNtAGIP1:: GUS fusions ( Fig 1A).…”

Section: Adding 35s Enhancer To Ntagip1 Broadens the Tissue Specificimentioning

confidence: 99%

A 100 bp GAGA motif-containing sequence in AGAMOUS second intron is able to suppress the activity of CaMV35S enhancer in vegetative tissues

Liu¹,

Zou

Wang³

et al. 2020

PLoS ONE

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“…The full 343 bp segment was therefore designated the 'CaMV 35S promoter', while the 46 bp segment was considered as the so-called 'minimal promoter' 29 . In follow-up studies, these 343 bp could then be further subdivided into several individual stretches, which would promote expression in different cell types or tissues, in either an additive or combinatorial fashion 30,31 . Based on these groundbreaking findings, numerous versions of the promoter emerged over the course of the following years; for example, simply placing two CaMV35S promoters in a tandem led to enhanced strength of the expression system 32 .…”

Section: The Camv 35s Promoter (1985-2000)mentioning

confidence: 99%

A short history of the CaMV 35S promoter

Somssich

2018

Preprint

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In an organism, be it plant, animal or human, almost every gene has its own promoter sequence, which is typified as a DNA stretch that controls how a gene is expressed in a cell. Hence, the activity of a promoter controls in which cell type, during which developmental stage or during what environmental condition a certain gene is expressed. However, the most widely used promoter in plant biotechnology is actually not derived from a plant, but a pathogenic virus. How and why did that happen? Here's a short history of the CaMV 35S promoter.

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“…The 90-35S promoter, containing a normal TATA-box and an ASF1 transcription factor-binding site (30), displayed a very low level of promoter activity in transgenic tobacco leaves. The transformants harboring the N1 construct, containing the sequence between –1415 and –115 of the PNZIP promoter, exhibited high GUS activity, not only in leaves but also in roots.…”

Section: Resultsmentioning

confidence: 99%

Tissue-specific expression of the PNZIP promoter is mediated by combinatorial interaction of different cis-elements and a novel transcriptional factor

Yang

et al. 2009

Nucleic Acids Research

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Recent studies demonstrated that PNZIP and its homologs encode a special cyclase and play an important role in chlorophyll biosynthesis in higher plants. To investigate the molecular mechanism governing the PNZIP gene, the PNZIP promoter was isolated and analyzed. Deletion analysis indicated that G-box is an important element in the regulation of the reporter gene expression. Further mutation assay demonstrated that G-box and GATACT elements are necessary and sufficient for the high and tissue-specific expression of the GUS gene. Using yeast one-hybrid screening, we have isolated a novel tobacco bZIP protein, NtbZIP, which can specifically recognize the G-box of the PNZIP promoter. The NtbZIP protein shares a limited amino acid homology to Arabidopsis ABI5 and AtAREB1 and very low homology to other bZIP proteins. Northern blot analysis showed that the NtbZIP gene is not induced by exogenous ABA and is expressed in different tobacco organs. Cotransformation assays showed that the NtbZIP protein could activate the transcription of the GUS gene driven by the PNZIP promoter. Transgenic tobaccos analysis demonstrated that constitutively expressing antisense NtbZIP gene resulted in a lower NTZIP synthesis and reduced chlorophyll levels. We suggest that NTZIP is a target gene of NtbZIP, which is involved in the regulation of chlorophyll biosynthesis.

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Multiple cis regulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants.

Cited by 255 publications

References 36 publications

A 100 bp GAGA motif-containing sequence in AGAMOUS second intron is able to suppress the activity of CaMV35S enhancer in vegetative tissues

A 100 bp GAGA motif-containing sequence in AGAMOUS second intron is able to suppress the activity of CaMV35S enhancer in vegetative tissues

A short history of the CaMV 35S promoter

Tissue-specific expression of the PNZIP promoter is mediated by combinatorial interaction of different cis-elements and a novel transcriptional factor

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