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Identification of a novel ovine LH-beta promoter region, which dramatically enhances its promoter activity.

Aherrahrou R, Aherrahrou Z, Erdmann J, Moumni M - Springerplus (2015)

Bottom Line: An increase in the promoter activity of the whole region was demonstrated compared to the empty vector.More interestingly, the unpublished region significantly enhanced the promoter activity compared to the known region alone.Taken together, a novel region was identified and confirmed in sheep which contained a promoter activity rich with binding sites for a putative regulatory element as shown in silico.

View Article: PubMed Central - PubMed

Affiliation: Institute for Integrative and Experimental Genomics (IIEG), Luebeck University, Luebeck, Maria-Goeppert-Str. 1, 23562 Lübeck, Germany ; Department of Biology, Faculty of Sciences, Moulay Ismail University, Zitoune, BP 11201, 50000 Meknes, Morocco.

ABSTRACT
The luteinizing hormone beta subunit (LH-beta) gene plays a critical role in reproduction. In order to characterize and analyze the promoter region of LH-beta in sheep, a genomic library was constructed in phage lambda gt 10 and screened. A novel region of 1,224 bp upstream from the targeted LH-beta gene was identified. Blasting this sequence showed a perfect homology for the first 721 bp sequence with an upstream ovine LH-beta sequence in the database. However, the remaining 5'-503 bp showed no sequence matching. DNA from Moroccan breeds was isolated and the whole region was amplified and confirmed by sequencing. To further confirm the promoter activity of this region, an in vitro analysis using a luciferase assay was carried out. An increase in the promoter activity of the whole region was demonstrated compared to the empty vector. More interestingly, the unpublished region significantly enhanced the promoter activity compared to the known region alone. To predict putative transcription factor binding-sites (TFBSs), an in silico analysis was performed using the TFSEARCH program. The region features many TFBSs and contains two palindrome sequences of 17- and 18-bp. Taken together, a novel region was identified and confirmed in sheep which contained a promoter activity rich with binding sites for a putative regulatory element as shown in silico.

No MeSH data available.


In-silico transcription factor binding sites (TFBS) analysis of the whole promoter region of the LH beta gene. The in silico analysis of the promoter region was carried out using the TFSEARCH software program. The localization of each TF is shown in the figure according to its position from the ATG start codon. The first (unpublished) part of the promoter is shown.
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Fig3: In-silico transcription factor binding sites (TFBS) analysis of the whole promoter region of the LH beta gene. The in silico analysis of the promoter region was carried out using the TFSEARCH software program. The localization of each TF is shown in the figure according to its position from the ATG start codon. The first (unpublished) part of the promoter is shown.

Mentions: The TFSEARCH in silico program was used to predict potential regulatory elements within the whole region containing previously and newly identified regions. The analysis of the whole region revealed more than 30 putative regulatory elements (Figs. 3, 4). In addition, two extended perfect palindrome sequences of 17- and 18-bp were also identified. Palindromic sequences are thought to be regulatory sites for steroid hormones (Aumais et al. 1996).Fig. 3


Identification of a novel ovine LH-beta promoter region, which dramatically enhances its promoter activity.

Aherrahrou R, Aherrahrou Z, Erdmann J, Moumni M - Springerplus (2015)

In-silico transcription factor binding sites (TFBS) analysis of the whole promoter region of the LH beta gene. The in silico analysis of the promoter region was carried out using the TFSEARCH software program. The localization of each TF is shown in the figure according to its position from the ATG start codon. The first (unpublished) part of the promoter is shown.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4554545&req=5

Fig3: In-silico transcription factor binding sites (TFBS) analysis of the whole promoter region of the LH beta gene. The in silico analysis of the promoter region was carried out using the TFSEARCH software program. The localization of each TF is shown in the figure according to its position from the ATG start codon. The first (unpublished) part of the promoter is shown.
Mentions: The TFSEARCH in silico program was used to predict potential regulatory elements within the whole region containing previously and newly identified regions. The analysis of the whole region revealed more than 30 putative regulatory elements (Figs. 3, 4). In addition, two extended perfect palindrome sequences of 17- and 18-bp were also identified. Palindromic sequences are thought to be regulatory sites for steroid hormones (Aumais et al. 1996).Fig. 3

Bottom Line: An increase in the promoter activity of the whole region was demonstrated compared to the empty vector.More interestingly, the unpublished region significantly enhanced the promoter activity compared to the known region alone.Taken together, a novel region was identified and confirmed in sheep which contained a promoter activity rich with binding sites for a putative regulatory element as shown in silico.

View Article: PubMed Central - PubMed

Affiliation: Institute for Integrative and Experimental Genomics (IIEG), Luebeck University, Luebeck, Maria-Goeppert-Str. 1, 23562 Lübeck, Germany ; Department of Biology, Faculty of Sciences, Moulay Ismail University, Zitoune, BP 11201, 50000 Meknes, Morocco.

ABSTRACT
The luteinizing hormone beta subunit (LH-beta) gene plays a critical role in reproduction. In order to characterize and analyze the promoter region of LH-beta in sheep, a genomic library was constructed in phage lambda gt 10 and screened. A novel region of 1,224 bp upstream from the targeted LH-beta gene was identified. Blasting this sequence showed a perfect homology for the first 721 bp sequence with an upstream ovine LH-beta sequence in the database. However, the remaining 5'-503 bp showed no sequence matching. DNA from Moroccan breeds was isolated and the whole region was amplified and confirmed by sequencing. To further confirm the promoter activity of this region, an in vitro analysis using a luciferase assay was carried out. An increase in the promoter activity of the whole region was demonstrated compared to the empty vector. More interestingly, the unpublished region significantly enhanced the promoter activity compared to the known region alone. To predict putative transcription factor binding-sites (TFBSs), an in silico analysis was performed using the TFSEARCH program. The region features many TFBSs and contains two palindrome sequences of 17- and 18-bp. Taken together, a novel region was identified and confirmed in sheep which contained a promoter activity rich with binding sites for a putative regulatory element as shown in silico.

No MeSH data available.