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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in the promoter activity of the whole region was demonstrated compared to the empty vector.More interestingly, the unpublished region significantly enhanced the promoter activity compared to the known region alone.Taken together, a novel region was identified and confirmed in sheep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 More interestingly, the unpublished region significantly enhanced the promoter activity compared to the known region alone.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.


Schematic representation of the promoter region of the LH beta gene from a library screening of the Moroccan sheep database sequence: Homology sequence analysis revealed that the two sequences shared more than 96% homology with the reference sequence available in the database after alignment (Brown et al. 1993).
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Fig1: Schematic representation of the promoter region of the LH beta gene from a library screening of the Moroccan sheep database sequence: Homology sequence analysis revealed that the two sequences shared more than 96% homology with the reference sequence available in the database after alignment (Brown et al. 1993).

Mentions: The LH-beta gene was cloned from a sheep genomic library, which was constructed in phage lambda gt 10 and screened. Six clones were selected and the corresponding DNA products were sequenced and analyzed using a Blast program. The analysis of the identified clone showed a perfect homology with the sequence of ovine LH-beta published in the database (http://blast.ncbi.nlm.nih.gov, Brown et al. 1993). However, our identified sequence contained an additional promoter fragment of 503 bp (in the 5′ side), which has not yet been published (Fig. 1 and Additional file 1: Figure S1 in the supporting information).Fig. 1


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)

Schematic representation of the promoter region of the LH beta gene from a library screening of the Moroccan sheep database sequence: Homology sequence analysis revealed that the two sequences shared more than 96% homology with the reference sequence available in the database after alignment (Brown et al. 1993).
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig1: Schematic representation of the promoter region of the LH beta gene from a library screening of the Moroccan sheep database sequence: Homology sequence analysis revealed that the two sequences shared more than 96% homology with the reference sequence available in the database after alignment (Brown et al. 1993).
Mentions: The LH-beta gene was cloned from a sheep genomic library, which was constructed in phage lambda gt 10 and screened. Six clones were selected and the corresponding DNA products were sequenced and analyzed using a Blast program. The analysis of the identified clone showed a perfect homology with the sequence of ovine LH-beta published in the database (http://blast.ncbi.nlm.nih.gov, Brown et al. 1993). However, our identified sequence contained an additional promoter fragment of 503 bp (in the 5′ side), which has not yet been published (Fig. 1 and Additional file 1: Figure S1 in the supporting information).Fig. 1

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.