scholarly journals G1 point mutation in growth differentiation factor 9 gene affects litter size in Sudanese desert sheep

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 ◽  
...  
Keyword(s):  
Point Mutation ◽  
Litter Size ◽  
Reference Sequence ◽  
Wild Type ◽  
Mutant Type ◽  
Type Allele ◽  
Sheep Breeds ◽  
Differentiation Factor ◽  
Growth Differentiation Factor 9 ◽  
Gdf9 Gene

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.

scholarly journals Polymorphism and Bioinformatics Analysis of Growth Differentiation Factor 9 Gene in Lori Sheep

2015 ◽  
Vol 15 (2) ◽  
pp. 337-348 ◽  
Author(s):  
Pouya Zamani ◽  
Ramin Abdoli ◽  
Ali Deljou ◽  
Hosein Rezvan
Keyword(s):  
Litter Size ◽  
Major Gene ◽  
Protein Structures ◽  
Single Strand Conformational Polymorphism ◽  
Differentiation Factor ◽  
Growth Differentiation Factor 9 ◽  
Exon 1 ◽  
Isoelectric Ph ◽  
Gene Exon ◽  
Gdf9 Gene

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.

Genetic polymorphism in fecundity gene (FecG) among Indiansheep breeds Balangir, Shahabadi and Bonpala

2016 ◽  
Author(s):  
Jowel Debnath ◽  
Ran Vir Singh
Keyword(s):  
Litter Size ◽  
Transforming Growth Factor ◽  
Natural Habitat ◽  
Factor B ◽  
Sheep Breeds ◽  
Homozygous Genotype ◽  
Average Litter Size ◽  
Pcr Rflp ◽  
Selection Of ◽  
Gdf9 Gene

FecG (GDF9) is a member of the transforming growth factor-b (TGF- b) superfamily, have been shown to be essential for follicular growth and ovulation. Different mutations in FecG gene caused increased ovulation and infertility in sheep. The present study was designed for screening polymorphism of FecG gene in 250 selected ewes from different sheep flocks representing Balangir (100), Shahabadi (100) and Bonpala (50) by employing forced PCR-RFLP technique. Genomic DNA was extracted from the blood of Balangir, Shahabadi and Bonpala matured ewes with average litter size varying from 1.00± 0.00 to 1.14±0.02 at different parities. Digestion of FecG (GDF9) gene with DdeI restriction enzyme resulted into FecGHH homozygous genotype. In all three sheep breeds, genotypic frequencies of FecGHH were 100% and gene frequency of H allele was unity. This indicates that the FecG gene is fixed in the Balangir, Shahabadi and Bonpala population in the natural habitat. Litter size of Balangir and Bonpala sheep breeds were single but in Shahabadi sheep twin was recorded. In present study all the animals of three breeds were homozygous for FecG and there was no infertility observed in above mentioned breeds in field condition and organized farm, which is not in consonance with previous report. The observed effects could be caused by linkage disequilibrium with other nearby loci. The study revealed that FecG gene is not a reliable genetic marker for selection of high prolificacy in sheep.

scholarly journals Fingerprinting of fecB gene in five Egyptian sheep breeds

2009 ◽  
Vol 25 (3-4) ◽  
pp. 205-212 ◽  
Author(s):  
Hanafy el ◽  
Saadani el
Keyword(s):  
Point Mutation ◽  
Restriction Enzyme ◽  
Base Pair ◽  
Genomic Dna ◽  
Restriction Site ◽  
Wild Type ◽  
Specific Primer ◽  
Sheep Breeds ◽  
Pcr Rflp ◽  
Pcr Products

Recently, many aspects of FecB gene, including reproductive endocrinology, organs development and body mass have been studied. FecB has an additive effect on litter size and ovulation. The present investigation was carried out to study polymorphism by forced PCR-RFLP of FecB gene in five Egyptian local sheep breeds and its comparison with other foreign sheep breeds. Genomic DNA was isolated from a total of 100 animals of Egyptian sheep breeds namely Rahmani, Ossimi, Awassi, Barki and Awassi x Barki crossbred. Forced PCR of the FecB gene 190 base pair (bp) was amplified using specific primer designed to introduc a point mutation in the resulting PCR products with FecB carrier sheep containing an AvaII restriction site (G|GACC), whereas products from noncarriers lacked (of) this site. Digestion of FecB gene 190 base pair with AvaII restriction enzyme resulted in non carrier 190 bp band (wild type) in all the animals belonging to the five Egyptian breeds studied revealing absence of this restriction site in those five breeds. .

scholarly journals Variant <i>GDF9</i> mRNA is likely not the main cause of larger litter size in Iranian Lori-Bakhtyari, Shal, Ghezel, and Afshari sheep breeds

2017 ◽  
Vol 60 (2) ◽  
pp. 119-129 ◽  
Author(s):  
Shahin Eghbalsaied ◽  
Farzad Rashidi Khorasgani ◽  
Hamid-Reza Amini ◽  
Majid Farahi ◽  
Maryam Davari ◽  
...  
Keyword(s):  
Litter Size ◽  
Signal Peptide ◽  
Large Dataset ◽  
Sheep Breeds ◽  
Signal Peptide Region ◽  
Twin Birth ◽  
Significant Difference ◽  
Pcr Rflp ◽  
Peptide Region ◽  
Gdf9 Gene

Abstract. This study was carried out to screen the GDF9 gene and evaluate the polymorphism effect on litter size of four Iranian sheep breeds using the PCR-RFLP and PCR-SSCP methods. First, sequencing of the GDF9 gene in 16 twin-birth, 4 triplet-birth, and 2 infertile ewes showed that, in addition to G2, G3, G4, G5, and G6 mutations that have been previously reported in other breeds, a new G0 mutation, called C25T, exists in the GDF9 sequence of 1 out of 22 ewes and causes L9F substitution in the signal peptide region. None of the triplet-birth or infertile ewes carried G1, G4, G7, FecGE, G8, or FecGT mutations. In the second experiment, a large dataset was used: 605 individuals including 496 ewes (145 Afshari, 54 Shal, 126 Ghezel, and 171 Lori-Bakhtyari sheep), and 109 rams (26 Afshari, 23 Shal, 10 Ghezel, and 50 Lori-Bakhtyari sheep. There were no sheep carrying the G7, G8, or Thoka mutations. Among all 109 rams that were used in this study, none of them were homozygous for the G1 mutation. Moreover, abundance of heterozygote rams (G1/G+) varied from 0.0 (Afshari) to 28.6 % (Lori-Bakhtyari and Ghezel). The highest and the lowest frequencies of the G4 mutation were 30.6 and 3.0 % in Shal and Afshari breeds, respectively. Moreover, G4 abundance varied from 0.0 to 42.3 %, from 3.0 to 26.9, and from 3.0 to 30.6 % in rams, ewes, and overall, respectively. There was a significant difference in the abundance of G1 and G4 mutations between breeds. However, neither the G1 nor the G4 mutation was associated with litter size in Afshari, Ghezel, Lori-Bakhtyari, or Shal sheep breeds. In conclusion, the results of this study showed that GDF9 G1 and G4 mutations are not the reason for higher litter size in Iranian sheep. Moreover, the GDF9 G0 and G6 mutations do not cause triplet births or infertility in Iranian ewes. Therefore, it is unlikely that variant GDF9 mRNA induces larger litter size or infertility in Iranian ewes.

scholarly journals MULTIPLEX PCR BASED SINGLE NUCLEOTIDE POLYMORPHISMS ANALYSIS OF HBS POINT MUTATION

2020 ◽  
pp. 1-4
Author(s):  
Jignisha S Patel ◽  
Jignaben P Naik ◽  
Yazdi M Italia
Keyword(s):  
Multiplex Pcr ◽  
Point Mutation ◽  
Sickle Cell ◽  
Mutant Allele ◽  
Snp Analysis ◽  
Wild Type Allele ◽  
Wild Type ◽  
Single Nucleotide ◽  
Type Allele ◽  
Cell Carrier

Introduction: Sickle hemoglobin (HbS), an autosomal recessive hemoglobinopathy cause of Sickle cell disease (SCD), is widely sprayed around the globe affecting millions of people . SCD results from single nucleotide polymorphism (SNP) or point mutation causing amino acid substitution from Glutamic acid to Valine leads to sickled shape red blood cells. SNPs can be well studied by using allele-specific amplification (ASA) technique. Aims & Objective: To develop a simple, rapid, easy and accurate genotyping method for SNP analysis of SCD. Materials and methods: By performing different tests, a well characterized sample panel of 150 different types of samples was prepared. From this sample panel DNA was extracted and used for SNP-genotyping of SCD. Specific primers were used for performing monoplex PCR amplification of wild type allele (HbAA) and mutant allele (HbSS) were performed individually. By using the same primers multiplex PCR assay was experimented. Results and conclusion: This is a simple and low cost molecular method for the detection of point mutation and useful tool for the diagnosis of SCD. The entire analysis can be performed in one reaction mixture, which results in higher speed, higher accuracy, and the need for smaller samples. This technique might be of great value for genotyping of homozygous sickle cell patients (SS) and heterozygous sickle cell trait (AS). But we found one discrepancy with double heterozygous (sickle β-thalassaemia) samples. We were not able to differentiate sickle cell carrier state (AS) from the double heterozygous like sickle β-thalassaemia state. So we conclude that for simultaneous detection of thalassaemia along with sickle cell requires addition of more primers specific for thalassaemia mutation. In addition to this when two bands, one for wild type allele and second for mutant allele appears, care must be taken to conclude whether the person is a sickle cell carrier (AS) or having double heterozygous (sickle β-thalassaemia) like condition.

A novel genotyping technique for discriminating LVAS-associated hotspot mutations in SLC26A4 gene

2020 ◽  
Author(s):  
Chen Zhou ◽  
Xiangman Zou ◽  
Cuiying Peng ◽  
Guoqiang Gao ◽  
Zifen Guo
Keyword(s):  
Ct Scan ◽  
Primer Extension ◽  
Wild Type Allele ◽  
Wild Type ◽  
Specific Primers ◽  
Mutant Type ◽  
Type Allele ◽  
Slc26a4 Gene ◽  
Allele Specific ◽  
Hotspot Mutations

Abstract An increasing number of biological and epidemiological evidence suggests that IVS7-2A>G and 2168A>G mutations of solute carrier family 26, member 4 ( SLC26A4 ) gene plays a critical role in the development of large vestibular aqueduct syndrome (LVAS). In this study, we developed a rapid genotyping method for discriminating LVAS-associated hotspot mutations in SLC26A4 gene. The genotyping technique consists of 3’ terminal exonuclease-resistant phosphorothioate-modified allele specific primer extension mediated by exo + polymerase. In PCR amplification by Pfu polymerase, allelic specific primers perfectly matching wild type allele were extended while no specific products were yielded from primers targeting mutant type allele. Similarly, allelic specific primers perfectly matching mutant type allele were extended and no specific products were observed from primers targeting wild type allele . The clinical application of 3’ terminal phosphorothioate-modified allele specific primers extension mediated by Pfu polymerase identified both homozygous for SLC26A4 gene IVS7-2A>G mutation in two LVAS patients diagnosed as by temporal bone CT scan. The genetic results from this method is consistent with that of DNA sequencing. The data suggest that exo + polymerase-mediated 3’ terminal phosphorothioate-modified primer extension is reliable in the identification of SLC26A4 gene hotspot mutation prior to high resolution CT scan. The method is extremely suitable for quickly molecular etiologic screening and early diagnosis and aggressive prevention therapy of LVAS.

scholarly journals Genetic Effects of Single Nucleotide Polymorphisms in the Goat GDF9 Gene on Prolificacy: True or False Positive?

Animals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 886 ◽  
Author(s):  
Xinyu Wang ◽  
Qing Yang ◽  
Sihuan Zhang ◽  
Xiaoyu Zhang ◽  
Chuanying Pan ◽  
...  
Keyword(s):  
Single Nucleotide Polymorphisms ◽  
Litter Size ◽  
Information Search ◽  
Reproductive Traits ◽  
High Fertility ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
Growth Differentiation Factor 9 ◽  
Linkage Effects ◽  
Gdf9 Gene

Goat reproductive traits are complex quantitative traits controlled by polygenes and multipoint. To date, some high-fertility candidate genes in livestock have been unearthed and the growth differentiation factor 9 (GDF9) gene is one of them, which plays a crucial role in early folliculogenesis. According to the relevant previous studies and the National Center for Biotechnology Information Search database (NCBI), a total of 45 single nucleotide polymorphisms (SNPs) have been detected in the goat GDF9 gene, but which one or which ones have important effects on goat fecundity is still uncertain. Hence, in order to find effective molecular markers for goat genetic breeding and accelerate the goat improvement, this study summarized and classified the above 45 SNPs into four kinds, as well as compared and analyzed the same SNP effects and the different SNPs linkage effects on the reproductive traits in different goat breeds. Since there were many SNPs in the goat GDF9 gene, only 15 SNPs have been identified in more than 30 goat breeds worldwide and they showed different effects on the litter size. Therefore, this study mainly chose these 15 SNPs and discussed their relationship with goat productivity. Results showed that three non-synonymous SNPs A240V, Q320P, and V397I and three synonymous ones L61L, N121N, and L141L played a “true” role in the litter size trait in many goat breeds around the world. However, the regulatory mechanisms still need further research. These results provide an effective tool for follow-up research developing the goat molecular breeding strategies and improving the goat reproductive traits.

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