GDF9

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1 - بررسی چند شکلی ژن GDF9 در گوسفند قره گل و ارتباط آن با چند قلوزایی
رضا سید شریفی نجات بادبرین امجد بهمنی علی مجتهدین

زمینه و هدف:یکی از صفات بسیار مهم و اقتصادی در پرورش گوسفند چند قلوزایی است. مطالعات ژنتیکی مشخص نموده است که چند قلوزایی تحت تاثیر مجموعهای از ژنها معروف به ژنهای باروری قرار دارد. از جمله این ژن ها، ژن GDF9است که بر روی کروموزوم شماره 5 گوسفند شناسایی گردید. پژوهش چکیده کامل

زمینه و هدف:یکی از صفات بسیار مهم و اقتصادی در پرورش گوسفند چند قلوزایی است. مطالعات ژنتیکی مشخص نموده است که چند قلوزایی تحت تاثیر مجموعهای از ژنها معروف به ژنهای باروری قرار دارد. از جمله این ژن ها، ژن GDF9است که بر روی کروموزوم شماره 5 گوسفند شناسایی گردید. پژوهش حاضر با هدف بررسی چند شکلی ژن GDF9و رابطه آن با صفت چندقلوزایی در گوسفند قره گل انجام گرفت. روش کار:برای این منظور از 100 راس گوسفند ماده قره گل در ایستگاه اصلاح نژاد قره گل سرخس به صورت تصادفی نمونه خون تهیه و اطلاعات زایش آن ها ثبت گردید. پس از استخراج DNAبا استفاده از یک جفت آغازگر اختصاصی یک قطعه 1503 جفت بازی از ژن GDF9تکثیر شد. قطعه تکثیر شده با استفاده از آنزیم Rsa Iهضم گردید. با استفاده از نرم افزار SAS 9.1ارتباط بین ژنوتیپهای شناسایی شده و صفت چند قلوزایی بررسی شد.یافته ها:نتایج حاصل از هضم آنزیمی وجود جهش را در این جایگاه ژنی تایید نمود و رابطه معنیدار(05/0P) بین ژنوتیپهای مشاهده شده و صفت چند قلوزایی نشان داد، به طوری که حیوانات هتروزیگوت نسبت به هموزیگوتها میزان چند قلوزایی بالاتری داشتند.نتیجه گیری: جهش ایجاد شده در ژن GDF9جهش مفیدی بوده و اقدام برای تثبیت این جهش در طی سالهای آینده به احتمال زیاد موجب افزایش چند قلوزایی، افزایش راندمان تولید مثلی و در نهایت افزایش درآمد دامداران خواهد شد. پرونده مقاله

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2 - Novel Single Nucleotide Polymorphisms (SNPs) in Two Oogenesis Specific Genes (BMP15, GDF9) and Their Association with Litter Size in Markhoz Goat (Iranian Angora)
ب. عارف‌نژاد ی. مهدی‌زاده آ. جوانمرد م.ج. ضمیری ع. نیازی

BMP15 and GDF9 are two oogenesis specific genes play a pivotal role in female fertility in mammals and potential for improvement of prolificacy in marker-assisted selection. The aim of present research was to investigate the variation and association between BMP15 and G چکیده کامل

BMP15 and GDF9 are two oogenesis specific genes play a pivotal role in female fertility in mammals and potential for improvement of prolificacy in marker-assisted selection. The aim of present research was to investigate the variation and association between BMP15 and GDF9 polymorphism and litter size in Markhoz goats. The sequence variability of the different amplified fragments utilized for genotyping was further investigated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and sequencing. Sequencing technology allowed the detection of two nucleotide variations: A both non-functional mutation (G/A) transition at nt 1026 known as P3) and a synonymous mutation at 1049 position which for the first time was found at the BMP15 gene. Also, GDF9 gene exhibited a polymorphism on BcnI site (1159 bp) within exon 2 of amplified PCR products (glutamine/proline). Our analysis using general linear model (GLM) statistical methods showed litter size was significantly influenced by genotypes of GDF9 (P<0.01). Heterozygous genotype showed higher litter size than homozygous genotypes (P<0.01). The conformation of this founding would need large sample size and test to screen in other indigenous goats. پرونده مقاله

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3 - A Profile of Single Nucleotide Polymorphisms in Fecundity Genes Among Iranian Sheep Breeds by Using Polymerase Chain Reaction–Restriction Fragment Length Polymorphism (PCR-RFLP) Method
پ. پتکی س.ض. میرحسینی ف. افراز س.م.ف. وحیدی

Ovulation rate and litter size are traits of fecundity and have a very remarkable economic value that leads to an increase in the production of meat, wool and milk. Therefore, exploring on the candidate and functional genes for these traits is very important. Growth dif چکیده کامل

Ovulation rate and litter size are traits of fecundity and have a very remarkable economic value that leads to an increase in the production of meat, wool and milk. Therefore, exploring on the candidate and functional genes for these traits is very important. Growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15) and bone morphogenetic protein receptor-1B (BMPR1B) are the most important of these genes containing single nucleotide polymorphisms (SNPs) markers that are associated with high fertility in sheeps. The aim of this study was to determine the allelic and genotypic frequencies of seven of these SNPs including; G1, G4, G7 and G8 in GDF9 gene, B2 and B4 in BMP15 gene and FecB in BMPR1B gene among 532 sheeps from eleven of Iranian native breeds using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) method. For each mutation region, polymerase chain reactions and digestion reactions with specific endonuclease enzymes, were performed. The results showed that G1 mutation presents among ten breeds except Zandi, G4 mutation in all breeds, and the FecB mutation in two breeds; Kalakuei and Zandi. The G7, G8, B2 and B4 mutations were not observed in any of breeds, and all sheeps were homozygote wild type for these four positions. پرونده مقاله

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4 - Polymorphisms in GDF9 Gene and Its Relationship with Litter Size in Five Breeds of Black Goats
G.Q. Zhu Q.I. Wang Y.G. Kang Y.Z. Lv B.Y. Cao

In this study, we investigated the relationship between the genetic polymorphism of growth differentiation factor 9 (GDF9) genes and the litter size in 384 individuals of five breeds of black goats. Four pairs of primers were designed to detect single nucleotide polymor چکیده کامل

In this study, we investigated the relationship between the genetic polymorphism of growth differentiation factor 9 (GDF9) genes and the litter size in 384 individuals of five breeds of black goats. Four pairs of primers were designed to detect single nucleotide polymorphism of GDF9 gene in goats by PCR-SSCP. The least square was used to analyze the relation between different genotypes and the litter size. The results showed that the PCR products from primer pair 1 (P1) displayed polymorphisms in three genotypes (AA, AB and BB) in big foot (BF) and Jintang (JT) black goats. For primer pairs of P2, P3 and P4, there was no polymorphism. The sequencing results revealed that there was a single nucleotide mutation (A792→G) in exon 2 of GDF9 gene in BF and JT black goats, and this mutation resulted in an amino acid change: valine→isoleucine. In BF and JT black goats, the average litter size in the third parity was significantly higher in genotype AA than both genotypes of AB and BB while the average little size of genotype AB was higher than that of genotype BB in the same parity. GDF9 gene could be therefore considered as a candidate gene for marker-assisted selection of litter size trait in goats. پرونده مقاله

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5 - Simultaneous Presence of G1 and G4 Mutations in Growth Differentiation Factor 9 Gene of Iranian Sheep
S. Eghbalsaied H. Amini S. Shahmoradi M. M. Farahi

The fecundity in sheep has been much interested for producers due to its economical importance. Growth differentiation factor 9 (GDF9) is an autosomal gene with over dominance effect for twosingle nucleotide polymorphisms(SNPs), which resulted in sterility in homozygote چکیده کامل

The fecundity in sheep has been much interested for producers due to its economical importance. Growth differentiation factor 9 (GDF9) is an autosomal gene with over dominance effect for twosingle nucleotide polymorphisms(SNPs), which resulted in sterility in homozygote mutant ewes and additive effect for three SNPs, without showing sterility event. In the present study, DNA was extracted from whole blood of 19 Iranian Afshari and 8 Shal sheep breeds. The extracted DNA was multiplied several folds for G1, G4 and G8 sites in GDF9 gene through polymerase chain reaction(PCR) technique and was sent for sequencing. Results showed that G1-GDF9 mutation (arginine to histidine shift) was detected in Afshari breed, while G4-GDF9 mutation (glutamic acid to lysine shift) was detected in both Afshari and Shal breeds. The observed SNPs were recorded with very low frequency (<8%) in fertile ewes with twining birth, while infertile ewe did not carry the mutant allele. This might indicate an association between these mutations and twin-births in Iranian breeds. Furthermore, these mutations could not be the reason for ewe infertility. In consistent with the rare frequency of infertility in Iranian sheep, the investigated population lacked the G8 or Thoka mutations, which are the most effective mutations in GDF9 gene.This may be due to lack or rare frequency of this SNP in Iranian sheep flocks. In conclusion, G1- and G4-GDF9 SNPs are segregating with very low frequency in Iranian sheep flocks, so that no homozygous ewe for these SNPs was detected in the current study. The same phenomenon could be the case for other important mutations and the sample size should be very large to rule out presence of such mutations in Iranian sheep breeds. پرونده مقاله

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6 - Investigation of GDF9 and BMP15 Polymorphisms in Mehraban Sheep to Find the Missenses as Impact on Protein
ا. احمدی ف. افراز ر. طالبی ع. فرح‌آور س.م.ف. وحیدی

Utilization of fecundity genes such as GDF9 and BMP15 can help improve reproductive traits in sheep breeding programme. To evaluate effects of missense mutations on protein function, the polymorphisms of GDF9 and BMP15 genes were screened in twelve mehraban sheep using چکیده کامل

Utilization of fecundity genes such as GDF9 and BMP15 can help improve reproductive traits in sheep breeding programme. To evaluate effects of missense mutations on protein function, the polymorphisms of GDF9 and BMP15 genes were screened in twelve mehraban sheep using DNA sequencing, followed by protein structure modeling. Six single nucleotide polymorphism (SNPs) known as FecG mutations (G1-G6), were detected in exons 1 and 2 of GDF9 gene. Mutations of G1 (GDF9 exon 1 g.2118 G>A), G4 (GDF9 exon 2 g.3451 T>C) and G6 (GDF9 exon 2 g.3974 G>A) have shown amino acid substitution. None polymorphism was detected in exon 1 and exon 2 of BMP15 gene. Based on identified polymorphisms, individuals were classified into three haplotypes of wild haplotype (without mutation), haplotype A (simultaneous mutations of G1, G2, G3 and G4) and haplotype B (simultaneous mutations of G5 and G6). The 3D-structure of GDF9 protein in A and B haplotypes was rotated 90˚ and 45˚ than wild haplotype, respectively. The missenses G1/p.Arg87His, G4/p.Glu241Lys and G6/p.Val332Ile variants were benign. However both the missenses of G7/p.Val371Met and G8/p.Ser315Phe were probably damaging. Phylogenetic tree of GDF9 gene revealed that individuals with A and B haplotypes were distinct from wild haplotypes with bootstrapping values of 63 and 76, respectively. In conclusion, GDF9 protein in A and B haplotypes showed a higher performance than wild haplotype due to synergism effects of simultaneous mutations. These types of mutations with effect on turn and helix of GDF9 conservative regions showed physical and functional interaction with TGFβ proteins. پرونده مقاله

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7 - Genetic Polymorphism of GDF9 Gene in Iranian Moghani Sheep Breed
F. Ala Noshahr A. Rafat

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 fact چکیده کامل

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. پرونده مقاله

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