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Cloning and Characterization of 5' Flanking Regulatory Sequences of AhLEC1B Gene from Arachis Hypogaea L.

Tang G, Xu P, Liu W, Liu Z, Shan L - PLoS ONE (2015)

Bottom Line: We further characterized the GUS expression patterns in the transgenic Arabidopsis lines.Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity.Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

View Article: PubMed Central - PubMed

Affiliation: Bio-Tech Research Centre, Shandong Academy of Agricultural Sciences / Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan, 250100, China.

ABSTRACT
LEAFY COTYLEDON1 (LEC1) is a B subunit of Nuclear Factor Y (NF-YB) transcription factor that mainly accumulates during embryo development. We cloned the 5' flanking regulatory sequence of AhLEC1B gene, a homolog of Arabidopsis LEC1, and analyzed its regulatory elements using online software. To identify the crucial regulatory region, we generated a series of GUS expression frameworks driven by different length promoters with 5' terminal and/or 3' terminal deletion. We further characterized the GUS expression patterns in the transgenic Arabidopsis lines. Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity. Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

No MeSH data available.


The second round of PCR amplification products of 5′ flanking regulation regions of peanut AhLEC1B gene by chromosome walking.The arrow indicates the target band.
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pone.0139213.g002: The second round of PCR amplification products of 5′ flanking regulation regions of peanut AhLEC1B gene by chromosome walking.The arrow indicates the target band.

Mentions: To investigate the major regulatory regions or elements of AhLEC1B, we isolated the promoter using chromosomal walking. As a result, the 5' flanking fragment of 1289 bp in length including the promoter region (1235bp) and 5'UTR sequences (54bp) was obtained from the peanut DNA library LP (Fig 2)


Cloning and Characterization of 5' Flanking Regulatory Sequences of AhLEC1B Gene from Arachis Hypogaea L.

Tang G, Xu P, Liu W, Liu Z, Shan L - PLoS ONE (2015)

The second round of PCR amplification products of 5′ flanking regulation regions of peanut AhLEC1B gene by chromosome walking.The arrow indicates the target band.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591277&req=5

pone.0139213.g002: The second round of PCR amplification products of 5′ flanking regulation regions of peanut AhLEC1B gene by chromosome walking.The arrow indicates the target band.
Mentions: To investigate the major regulatory regions or elements of AhLEC1B, we isolated the promoter using chromosomal walking. As a result, the 5' flanking fragment of 1289 bp in length including the promoter region (1235bp) and 5'UTR sequences (54bp) was obtained from the peanut DNA library LP (Fig 2)

Bottom Line: We further characterized the GUS expression patterns in the transgenic Arabidopsis lines.Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity.Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

View Article: PubMed Central - PubMed

Affiliation: Bio-Tech Research Centre, Shandong Academy of Agricultural Sciences / Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Jinan, 250100, China.

ABSTRACT
LEAFY COTYLEDON1 (LEC1) is a B subunit of Nuclear Factor Y (NF-YB) transcription factor that mainly accumulates during embryo development. We cloned the 5' flanking regulatory sequence of AhLEC1B gene, a homolog of Arabidopsis LEC1, and analyzed its regulatory elements using online software. To identify the crucial regulatory region, we generated a series of GUS expression frameworks driven by different length promoters with 5' terminal and/or 3' terminal deletion. We further characterized the GUS expression patterns in the transgenic Arabidopsis lines. Our results show that both the 65 bp proximal promoter region and the 52 bp 5' UTR of AhLEC1B contain the key motifs required for the essential promoting activity. Moreover, AhLEC1B is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding cis-regulatory elements.

No MeSH data available.