​POU1F1 Gene Polymorphism and its Associations with Body Weight of Osmanabadi Goat

Background: Osmanabadi is the premium meat type goat breed of Maharashtra, known for its adaptability and reproductive efficiency. Identification of superior germplasm and incorporating them into breeding programme is the need of hour for augmenting productivity of Osmanabadi as well as other indigenous goat breeds. Hence, the present research was aimed to study Pituitary Specific Transcription Factor I (POU1F1) gene polymorphism and its associations with body weight of Osmanabadi goat. Methods: Genomic DNA was extracted from 217 blood samples of randomly selected Osmanabadi goats. The 450 bp fragment of POU1F1 gene encompassing exon-6 and its flanking region (3’UTR) was PCR amplified. The PCR amplicon was subjected to RFLP using AluI and PstI restriction enzymes to identify polymorphism at nucleotide position 174 (T greater than C) in exon 6 and at nucleotide position 110 (T greater than C) in the 3’UTR of POU1F1 gene, respectively. The association of the observed allelic variants / genotypes with body weight of the animals was performed using One-way ANOVA. Result: The AluI PCR-RFLP of 450 bp amplicon of POU1F1 gene in Osmanabadi goats revealed polymorphism with three different genotypes viz., ‘CC’, ‘TT’ and ‘TC’. The observed frequencies for TT, TC and CC genotypes were 0.604, 0.355 and 0.041 respectively. The frequencies of T and C alleles were found to be 0.781 and 0.219, respectively. However, PstI PCR-RFLP revealed monomorphic 3’UTR of POU1F1 gene with single TT genotype. We observed significant differences (P less than 0.01) between various genotypes at AluI exon-6 locus of POU1F1 gene and body weights at birth, 3 and 6 months of Osmanabadi goat. Highest body weight was recorded for TT genotype followed by TC and CC genotypes in all three age groups. Our findings indicate that the AluI PCR-RFLP locus of the exon-6 of POU1F1 gene can be used as a potent molecular marker for selection of superior stock of Osmanabadi goats.

Genetic Variability in Exon 3 Region of POU1F1 Gene and its Effect on Body Weight and Milk Yield in Malabari Goats

Background: POU1F1 is known as a major candidate gene coding for pituitary-specific transcription factor-1 regulating the production of anterior pituitary hormones like GH, TSH, LH, FSH and prolactin in small ruminants. However, little is known about the genetic variability of caprine POU1F1 gene in Indian breeds. Hence, the present study aimed at the identification of single nucleotide polymorphisms in the exon 3 region of POU1F1 and their association with body weight and milk production in Malabari goats. Methods: The present study was conducted during 2016-2018 on 100 Malabari goats at the University Goat and Sheep Farm, Mannuthy. Genomic DNA was extracted using standard phenol-chloroform method. The POU1F1 exon 3 locus was PCR amplified using appropriate primers. The 234 bp PCR product was subjected to single-strand conformation polymorphism (SSCP) analysis on 12% PAGE gel and silver-stained. The different SSCP patterns were sequenced and homologous sequences identified using Blast algorithms. Allelic-genotypic frequencies and the observed-expected heterozygosity at the locus were estimated. Data on body weights at birth, three, six, nine months of age and parity-wise lactation milk yield were used to find the association of POU1F1 exon 3 variants with body weights and milk yield using one-way ANOVA in General Linear Models for fixed effects. Result: PCR-SSCP revealed GG, GA genotypes with frequencies of 0.44 and 0.56 and G and A alleles with frequencies of 0.72 and 0.28. The observed and expected heterozygosity for the locus were 0.5657 and 0.4077 respectively. Sequencing of the PCR products revealed a synonymous single nucleotide polymorphism (g. 42C>T). GG homozygotes had higher body weight while GA heterozygotes had higher milk yield (p less than 0.05). It was concluded that the polymorphic exon 3 locus significantly contributed to variation in pre-pubertal body weight and milk yield and hence may be employed in the marker-assisted selection programmes for Malabari goats.

Effect of POU1F1 Gene Haplotypes on Eights and Milk Production of Awassi Sheep

The aim of the study was to detect polymorphism in the POU1F1 gene in Iraqi Awassi sheep breed, as well as to establish if haplotype of POU1F1 gene could be associated with productive traits. This study was carried out at Al-Kafeel station, Karbala city during the period of 01/10/2017 until 01/08/2018. The study included 46 Awassi ewes with their 52 lambs. The laboratory analyses were conducted at the Laboratory of Molecular Genetics, College of Agriculture, University of Basrah. Results showed the successfulness of the PCR amplification process for all six examined fragments of the POU1F1 gene. Gel electrophoresis was conducted using agarose 2%, the product sizes were 637bp, 789bp, 999bp, 868bp, 1190 bp, and 469bp for the fragments P1, P2, P3, P4, P5, and P6 respectively. The analysis of the nitrogen bases sequences of the POU1F1 gene for the studied fragments showed a change in 12 different sites of the gene. These changes resulted in 7 haplotypes of H1-H7. The results showed significant influence (P<0.05) in haplotypes of the POU1F1 gene on birth weight . However, haplotypes showed no significant effect on lambs’ weights at weaning, six months and daily weight gains during all periods. Statistical analysis showed that different haplotypes of the POU1F1 gene did not influence on milk productive traits. The examined fragments of the POU1F1 Awassi gene have been submitted to Gene Bank under the accession numbers (LC469323 to LC469349).

Associations between genetic variants of the <i>POU1F1</i> gene and production traits in Saanen goats

This study was conducted to determine the polymorphisms of the POU1F1 gene and their relationships with milk yield and components, litter size, birth weight, and weaning weight in goats. For this purpose, a total of 108 Saanen goats from two different farms (Bornova and Manisa) were used as animal materials. Polymorphisms at the exon 6 and the 3′ flanking region of the POU1F1 gene were determined by using PCR-RFLP with PstI and AluI restriction enzymes and DNA sequencing analyses. Two alleles and three genotypes were identified by AluI or PstI digestions of the POU1F1 gene. The genotypes frequencies of TT, TC, and CC were 64.8 %, 31.5 % and 3.7 % for the PstI locus; 54.6 %, 31.5 % and 13.9 % for the AluI locus, respectively. T allele frequencies (0.56 and 0.88 for the AluI locus, 0.80 and 0.81 for the PstI locus, respectively) were predominant in both loci at the Bornova and Manisa farms. In terms of POU1F1-AluI and POU1F1-PstI loci, two populations were found to be in Hardy–Weinberg equilibrium. In the POU1F1-AluI locus, significant associations were found between genotypes and lactation milk yield and litter size. Similarly, a significant relationship between genotypes and birth weight in the POU1F1-PstI locus (p<0.05) was determined. The TC and CC genotypes were observed to be higher than the TT genotype for lactation milk yield and litter size at the POU1F1-AluI locus. Birth weight was found to be higher in animals that have the CC genotype at the POU1F1-PstI locus. In conclusion, the POU1F1 gene can be used as a molecular marker for economic features like reproduction, growth, milk content and yield in Saanen goats.

Relationship between SNPs of POU1F1 Gene and Litter Size and Growth Traits in Shaanbei White Cashmere Goats

POU (Pit-Oct-Unc) class 1 homeobox 1 (POU1F1, or Pit-1) is a transcription factor that directly regulates pituitary hormone-related genes, as well as affects the reproduction and growth in mammals. Thus, POU1F1 gene was investigated as a candidate gene for litter size and growth performance in goats. In the current study, using direct DNA sequencing, c.682G > T, c.723T > G and c.837T > C loci were genotyped in Shaanbei white cashmere (SBWC) goats (n = 609), but c.876 + 110T > C was monomorphic. Besides, the c.682G > T locus was first identified by HinfI (Haemophilus influenzae Rf) restriction endonuclease. Association analysis results showed that the c.682G > T, c.837T > C loci and diplotypes were significantly associated with goat litter size (p < 0.05). The positive genotypes were GT and TT for the two SNPs, respectively, and the optimal diplotype was H3H7 (GTTT-TTTT). On the other hand, the c.682G > T, c.723T > G and c.837T > C strongly affected growth traits and body measurement indexes in SBWC goats (p < 0.05). The positive genotypes or allele of these SNPs were GT, G and TT, respectively. Additionally, the goats with H3H7 diplotype also had a greater growth status than others (p < 0.05). Here, individuals with same genotype had both a better litter size and growth traits, showing a positive correlation between these economic traits. Meanwhile, the positive genotypes of four SNPs were combined to obtain the optimal diplotype, which was also H3H7. These SNPs, especially the diplotype, could be used for the genomic selection of excellent individuals with a greater litter size and better growth status in goat breeding.