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 More interestingly, the unpublished region significantly enhanced the promoter activity compared to the known region alone.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.Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-015-1182-5) contains supplementary material, which is available to authorized users.
Highlights
Sheep breeding in Morocco plays an important role in several areas of society, such as in traditional life and in the economy, and it represents the principle source for red meat
Identification of a novel promoter sequence region in the LH‐beta gene The luteinizing hormone beta subunit (LH-beta) gene was cloned from a sheep genomic library, which was constructed in phage lambda gt 10 and screened
Enhancement of the promoter activity by the novel identified region In order to test whether the unpublished novel promoter region is relevant for the promoter activity, fragments including either published, unpublished or the whole promoter region were amplified on genomic DNA from
Summary
Sheep breeding in Morocco plays an important role in several areas of society, such as in traditional life and in the economy, and it represents the principle source for red meat. In this study we focused on lutropin hormone, and more the beta subunit (LH-beta) which is secreted by the anterior pituitary (Sairam and Li 1975; Counis et al 1993; Moumni et al 1994; Lerrant et al 1995; Pelletier et al 1995). This hormone plays a determinant role in reproduction where it stimulates gonadal steroidogenesis and gametogenesis (Ma et al 2004).
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