Genetic Polymorphism of GDF9 Gene in Iranian Moghani Sheep Breed
محورهای موضوعی : Camel
1 - Department of Animal Science, University of Tabriz, Tabriz, Iran
2 - Department of Animal Science, University of Tabriz, Tabriz, Iran
کلید واژه: PCR, polymorphism, GDF9, Moghani sheep,
چکیده مقاله :
The families of TGF-β proteins are the most important growth factors in the ovary for growth and differentiation of early ovarian follicles. Three related oocyte-derived members of the transforming growth factor-β superfamily namely growth differentiation factor 9 (GDF9), BMP15 and BMPR-IB have been shown to be essential for follicular growth and ovulation. Different mutations in the GDF9 gene cause increased ovulation rate and infertility in a dosage-sensitive manner in sheep. In this study, blood samples were collected from 150 Moghani sheep breed using venojects treated with the anti-clot substance (EDTA) and subsequently their DNA content were salted out and extracted. The quantity and quality of extracted DNA was examined using spectrophotometery and gel electrophoresis, respectively. With using a pair of specific primers, the DNA fragment was amplified from exon 1 of GDF9 (462 bp). Digested PCR products with HhaI enzyme showed a G to A substiuation in GDF9 locus. The wild type allele of this gene (G/+) with two restriction site resulted DNA fragments of 156, 52 and 254 bp while the mutant allele (G/-) with one restriction site resulted two DNA fragments with the size of 52 and 410 bp. Genotype frequencies for G (+/+), G (+/-) and G (-/-) were 0.66, 0.24 and 0.1, respectively. From studied luci, GDF9 was polymorphic in Iranian Moghani sheep breed.
خانواده پروتئینهای TGF-B از فاکتورهای مهم رشد و تفرق فولیکولهای تخمدانی میباشند. سه ژن مهم از فوق خانواده B شامل فاکتور 9 (GDF9)، BMP15 و BMPR-IB در رشد و تخمکزایی ضروری میباشند. جهشهای مختلفی در ژن GDF9 سبب افزایش تخمکگذاری و تولید مثل در گوسفند میشود. در این مطالعه نمونههای خونی از 150 گوسفند نژاد مغانی جمعآوری و با افزودن EDTA نگهداری و به آزمایشگاه انتقال یافت. سپس DNA آنها به کمک روش نمکی استخراج گردید. کمیت و کیفیت DNA حاصله به ترتیب توسط روشهای اسپکتوفتومتری و ژل الکتروفورز بررسی شد. توسط یک جفت پرایمر قطعهای از DNA ( bp462) از اگزون ژن GDF9 استحصال گردید. محصولات حاصل از هضم PCR توسط آنزیم HhaI جانشینی باز G به A را در لوکاس GDF9 نشان داد. آلل وحشی این ژن (+/G) با دو جایگاه محدود کننده منجر به تولید سه قطعه DNA 156، 52 و bp 254 و آلل جهش یافته (-/G) با یک جایگاه برش سبب تولید دو قطعه 52 و bp 410 شد. فراوانی ژنوتیپی برای ژنوتیپهای (+/+)G، (-/+)G و (-/-)G به ترتیب 66/0، 24/0 و 1/0 به دست آمد. با توجه به مطالعه صورت گرفته چند شکلی ژن GDF9 در گوسفند نژاد مغانی معنیدار به دست آمد.
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