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Rapid validation of transcriptional enhancers using agrobacterium-mediated transient assay.

Identifieur interne : 000094 ( Main/Exploration ); précédent : 000093; suivant : 000095

Rapid validation of transcriptional enhancers using agrobacterium-mediated transient assay.

Auteurs : Yuan Lin [États-Unis] ; Fanli Meng [République populaire de Chine] ; Chao Fang [États-Unis] ; Bo Zhu ; Jiming Jiang [États-Unis]

Source :

RBID : pubmed:30873216

Abstract

Background

Enhancers are one of the most important classes of

Results

We developed a rapid enhancer validation system based on

Conclusions

In summary, we developed a rapid and efficient plant enhancer validation system based on a luciferase reporter and


DOI: 10.1186/s13007-019-0407-y
PubMed: 30873216
PubMed Central: PMC6402126


Affiliations:


Links toward previous steps (curation, corpus...)


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<b>Background</b>
</p>
<p>Enhancers are one of the most important classes of </p>
</div>
<div type="abstract" xml:lang="en">
<p>
<b>Results</b>
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<p>We developed a rapid enhancer validation system based on </p>
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<div type="abstract" xml:lang="en">
<p>
<b>Conclusions</b>
</p>
<p>In summary, we developed a rapid and efficient plant enhancer validation system based on a luciferase reporter and </p>
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<Year>2020</Year>
<Month>10</Month>
<Day>01</Day>
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<PubDate>
<Year>2019</Year>
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<Title>Plant methods</Title>
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<ArticleTitle>Rapid validation of transcriptional enhancers using agrobacterium-mediated transient assay.</ArticleTitle>
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<Abstract>
<AbstractText Label="Background" NlmCategory="UNASSIGNED">Enhancers are one of the most important classes of
<i>cis</i>
-regulatory elements (CREs) and play key roles in regulation of transcription in higher eukaryotes. Enhancers are difficult to identify because they lack positional constraints relative to their cognate genes. Excitingly, several recent studies showed that plant enhancers can be predicted based on their distinct features associated with open chromatin. However, experimental validation is necessary to confirm the predicted enhancer function.</AbstractText>
<AbstractText Label="Results" NlmCategory="UNASSIGNED">We developed a rapid enhancer validation system based on
<i>Nicotiana benthamiana</i>
. A set of 12 intergenic and intronic enhancers, identified in
<i>Arabidopsis thaliana</i>
, were cloned into a vector containing a minimal 35S promoter and a luciferase reporter gene, and were then infiltrated into
<i>N. benthamiana</i>
leaves mediated by agrobacterium. The enhancer activity of each candidate was quantitatively assayed based on bioluminescence measurement. The data from this luciferase-based validation was correlated with previous data derived from transgenic assays in
<i>A. thaliana</i>
. In addition, the relative strength of different enhancers for driving the reporter gene can be quantitatively compared. We demonstrate that this system can also be used to map the functional activity of a candidate enhancer under different environmental conditions.</AbstractText>
<AbstractText Label="Conclusions" NlmCategory="UNASSIGNED">In summary, we developed a rapid and efficient plant enhancer validation system based on a luciferase reporter and
<i>N. benthamiana</i>
-based leaf agroinfiltration. This system can be used for initial screening of leaf-specific enhancers and for validating candidate leaf enhancers from dicot species. It can potentially be used to examine the activity of candidate enhancers under different environmental conditions.</AbstractText>
</Abstract>
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<Keyword MajorTopicYN="N">Luciferase</Keyword>
<Keyword MajorTopicYN="N">Nicotiana benthamiana</Keyword>
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<name sortKey="Lin, Yuan" sort="Lin, Yuan" uniqKey="Lin Y" first="Yuan" last="Lin">Yuan Lin</name>
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