Detection of polymorphisms of growth differentiation factor 9 (GDF9) gene in Indonesian goats

2019 ◽  
Vol 3 (1) ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 104-112
Author(s):  
Amani Z. Abdelgadir ◽  
Lutfi M. A. Musa ◽  
Khaleel I. Jawasreh ◽  
Aubai. O. Saleem ◽  
Faisal El-Hag ◽  
...  

Background and Aim: Sudanese desert sheep encompass different sheep breeds named according to the different Sudanese tribes that rear them such as the Dubasi, Shugor, and Watish sheep. The objectives of this study were to screen for G1 point mutation in the polymorphic growth differentiation factor 9 (GDF9) gene, investigate its association with litter size, and construct the phylogeny of the different tribal breeds that belong to the Sudanese Desert sheep tribal types. Materials and Methods: Genomic DNA was extracted from whole blood of three tribal Desert sheep breeds (Dubasi, Watish, and Shugor) using the guanidine chloride method. Polymerase chain reaction-restriction fragment length polymorphism with HhaI restriction enzyme and sequencing techniques was used for genotyping the GDF9 locus for possible mutations associated with litter size in the three desert sheep tribal types. Results: G1 mutation in GDF9 caused the replacement of Arginine by Histidine at residue 87. The wild type allele (A) had the highest frequency, whereas the mutant type allele (a) had the lowest in all the sequenced subtypes. The genotype frequencies of the wild type ewes (AA) were higher than the heterozygous (Aa) and the mutant type (aa) frequencies in the three studied desert sheep types. No significant differences were found in the allele frequency between the three tribal types. Litter size was significantly influenced by the genotypes of GDF9 gene, parities, and subtypes (p≤0.01, 0.01, and 0.05, respectively). In the Watish sheep type, heterozygous sheep in their second parity recorded the highest litter size. Sequence alignment of GDF9 gene samples with the database entry indicated that all three tribal types were similar and identical to the reference sequence. The phylogenetic tree revealed that Shugor is the common ancestor of the studied types and Watish is more closely related to Shugor than Dubasi. This result mi ght partly explain the lower reproductive performance of Dubasi compared to Watish and Shugor. Conclusion: The presence of one copy of GDF9 gene increased litter size in the studied Sudanese Desert sheep. This locus may be used as a biomarker for litter size improvement through genotypic selection and allele or gene introgression.


2015 ◽  
Vol 15 (2) ◽  
pp. 337-348 ◽  
Author(s):  
Pouya Zamani ◽  
Ramin Abdoli ◽  
Ali Deljou ◽  
Hosein Rezvan

Abstract Growth differentiation factor 9 (GDF9) is a fecundity major gene affecting prolificacy in sheep. In the present study, genetic variation of a 380-bp fragment in GDF9 gene exon 1 was investigated in 100 Lori ewes. Single-strand conformational polymorphism (SSCP) and DNA sequencing methods were used to detect single nucleotide polymorphism (SNP) of the studied fragment. A SNP (g.306G>A), known as G1 mutation, with two genotypes (GG and AG) was found in two different SSCP patterns of GDF9 gene exon 1, deducing an amino acid (AA) exchange (p.Arg87His). Frequencies of the AG and GG genotypes were 37.65% and 62.35%, respectively. Also, the estimated allelic frequencies for the A and G alleles were 18.825% and 81.175%, respectively. The observed nucleotide sequences were subjected to alignment analysis and it was found that the studied fragment had more than 99.7% similarity with some sequences reported for other breeds of sheep. Two different secondary and 3D protein structures were predicted for A and G alleles. Moreover, the A and G alleles had different isoelectric pH values (8.7 and 9, respectively). The observed genotypes tended to have a significant association with litter size (P<0.10) where average litter size of GG ewes was slightly (20%) higher than for AG animals. With respect to the results of the present study, it seems that more studies are needed to evaluate the mutations in other fragments of this gene or other genes in Lori sheep.


Animals ◽  
2018 ◽  
Vol 8 (11) ◽  
pp. 200
Author(s):  
Zhangyuan Pan ◽  
Xiangyu Wang ◽  
Ran Di ◽  
Qiuyue Liu ◽  
Wenping Hu ◽  
...  

Growth differentiation factor 9 (GDF9) plays an important role in the early folliculogenesis of sheep. This study investigated the mRNA expression of ovine GDF9 in different tissues by real-time PCR. GDF9 exhibits significantly higher levels of expression (p < 0.01) in the ovary, relative to other tissues, indicating that its expression is tissue specific. To explore the regulatory mechanism of this tissue-specific expression, the methylation level of one CpG island (−1453 to −1854) of GDF9 promoter in ovary and heart was determined. In this region (−1987 to −1750), only the mC-4 site was present in the Sp4 binding site showed differential methylation between the heart and ovary; with increased (p < 0.01) methylation being observed in the heart. Additionally, the methylation level was negatively correlated with GDF9 mRNA expression (R = −0.75, p = 0.012), indicating that the methylation of this site plays an important role in transcriptional regulation of GDF9. The methylation effect of the mC-4 site was confirmed by using dual-luciferase. Site-directed mutation (methylation) of mC-4 site significantly reduced (p < 0.05) basal transcriptional activity of GDF9 promoter in oocytes. These results imply that methylation of GDF9 promoter CpG island mC-4 site may affect the binding of the Sp4 transcription factor to the GDF9 promoter region in sheep, thereby regulating GDF9 expression and resulting in a tissue-specific expression.


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