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ThesisItem Open Access MOLECULAR GENETIC STUDY OF POU1F1 GENE IN MANDYA AND NARI-SUWARNA SHEEP(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019-08) ABHILASH, H. R.; NAVEEN KUMAR, S.The present study was conducted to determine and compare the polymorphism of POU1F1 gene in Mandya and NARI-Suwarna Sheep. A total of 100 unrelated animals comprising of 50 each of Mandya and NARI-Suwarna sheep were utilized for the present study. Millers high salt method was followed to isolate the genomic DNA from the venous blood. The PCR amplification of exon 3 and exon 4 regions of POU1F1 gene was done by employing published primers, whereas, to amplify exon 6 region of POU1F1 gene primers were designed by using PRIMER 3 PLUS software. The PCR products of sizes 365 bp, 508 bp and 501 bp were resolved on 1.5 per cent agarose gel electrophoresis for exon 3, exon 4 and exon 6 region of POU1F1 gene, respectively. PCR-RFLP analysis of exon 3 and exon 4 regions of POU1F1 gene with AluI and EcoRI restriction enzymes revealed only one allele, A and one genotype AA in both Mandya and NARI-Suwarna sheep indicating monomorphism. The PCR-SSCP analysis of exon 6 region of POU1F1 gene revealed two patterns, P1 and P2. The frequency of P1 and P2 was 98 and 2 per cent, respectively in Mandya sheep and 90 and 10 per cent, respectively in NARISuwarna sheep. The alignment of P1 and P2 pattern sequences of Mandya sheep revealed two SNPs, G to C transversion at 109 bp position and T to A transversion at 112 bp position. Whereas, alignment of P1 and P2 pattern sequences of NARI-Suwarna sheep revealed 3 SNPs, T to G transversion at 218 bp, G to A transition at 225 bp and T to A transversion at 264 bp positions. From the above findings, it may be concluded that polymorphism was established at exon 6 region of POU1F1 gene in both Mandya and NARI-Suwarna sheep by PCR-SSCP analysis. However, present results need to be validated with large sample size.ThesisItem Open Access MOLECULAR GENETIC STUDY OF POU1F1 GENE IN MANDYA AND NARI-SUWARNA SHEEP(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019-08) ABHILASH, H. R.; NAVEEN KUMAR, S.The present study was conducted to determine and compare the polymorphism of POU1F1 gene in Mandya and NARI-Suwarna Sheep. A total of 100 unrelated animals comprising of 50 each of Mandya and NARI-Suwarna sheep were utilized for the present study. Millers high salt method was followed to isolate the genomic DNA from the venous blood. The PCR amplification of exon 3 and exon 4 regions of POU1F1 gene was done by employing published primers, whereas, to amplify exon 6 region of POU1F1 gene primers were designed by using PRIMER 3 PLUS software. The PCR products of sizes 365 bp, 508 bp and 501 bp were resolved on 1.5 per cent agarose gel electrophoresis for exon 3, exon 4 and exon 6 region of POU1F1 gene, respectively. PCR-RFLP analysis of exon 3 and exon 4 regions of POU1F1 gene with AluI and EcoRI restriction enzymes revealed only one allele, A and one genotype AA in both Mandya and NARI-Suwarna sheep indicating monomorphism. The PCR-SSCP analysis of exon 6 region of POU1F1 gene revealed two patterns, P1 and P2. The frequency of P1 and P2 was 98 and 2 per cent, respectively in Mandya sheep and 90 and 10 per cent, respectively in NARISuwarna sheep. The alignment of P1 and P2 pattern sequences of Mandya sheep revealed two SNPs, G to C transversion at 109 bp position and T to A transversion at 112 bp position. Whereas, alignment of P1 and P2 pattern sequences of NARI-Suwarna sheep revealed 3 SNPs, T to G transversion at 218 bp, G to A transition at 225 bp and T to A transversion at 264 bp positions. From the above findings, it may be concluded that polymorphism was established at exon 6 region of POU1F1 gene in both Mandya and NARI-Suwarna sheep by PCR-SSCP analysis. However, present results need to be validated with large sample size.
