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ThesisItem Open Access A COMPARATIVE STUDY ON GENETIC POLYMORPHISM OF MYOSTATIN AND FABP3 GENES IN MANDYA AND YALAGA SHEEP(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2021) MEENAKSHI, R. N.; R. NAGARAJA)The PCR-RFLP and SSCP polymorphisms of Myostatin (exon 1 and intron 1) and FABP3 (intron 3) genes were compared between Mandya and Yalaga sheep using the genomic DNA obtained from 50 each of Mandya and Yalaga sheep breeds. The Myostatin (497 bp and 414 bp) and FABP3 (355 bp) gene sequences were amplified by PCR employing published primers. PCR-RFLP analysis of Myostatin gene (497 bp) with DraI restriction enzyme revealed two genotypes, AB and BB with frequencies of 0.34 and 0.66 in Mandya sheep and 0.12 and 0.88 in Yalaga sheep, respectively. The allele frequencies for A and B were 0.17 and 0.83 in Mandya sheep and 0.06 and 0.94 in Yalaga sheep, respectively. PCR-RFLP analysis of FABP3 gene with BanII restriction enzyme revealed two genotypes, AA and AG with frequencies of 0.66 and 0.34 in Mandya sheep and 0.08 and 0.92 in Yalaga sheep, respectively. The gene frequencies for A and G were 0.83 and 0.17 in Mandya and 0.54 and 0.46 in Yalaga sheep, respectively. Multiple sequence alignment of B allele sequences of Myostatin (497 bp) gene and A allele sequences of FABP3 gene in Mandya and Yalaga sheep revealed T>C transition at 64 bp and C>T transition at 174 bp in Yalaga sheep. PCR-SSCP analysis of Myostatin (414 bp) gene revealed two genotypes, AA and AB with frequencies of 0.74 and 0.26 in Mandya sheep and 0.78 and 0.22 in Yalaga sheep, respectively. The gene frequencies for A and B were 0.87 and 0.13 and 0.89 and 0.11 in Mandya and Yalaga sheep, respectively. The alignment of A and B alleles of Myostatin gene in Mandya and Yalaga sheep revealed two SNPs: G>T transversion at position 46 bp and G>T transversion at position 287 bp in Yalaga sheep.ThesisItem Open Access COMPARATIVE GENETIC CHARACTERIZATION OF PROLACTIN GENE BETWEEN GIRIRAJA AND INDIGENOUS CHICKEN(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) AJEET SINGH; NAVEEN KUMAR, S.The present study was undertaken to determine and compare the genetic variations in prolactin gene among Giriraja and Indigenous chicken using PCR-RFLP technique. Blood samples were collected from 80 each of Giriraja and Indigenous chicken from Department of Poultry Science, Veterinary College, Bengaluru. The genomic DNA was isolated by following the conventional Phenol: Chloroform: Iso-amyl alcohol (25:24:1) method as described by Khosravinia et al. (2007). A 130/ 154 bp and 439 bp sequences of promoter regions 1 (24 bp indel at -358) and 2 (C-2402T and C-2161G) of prolactin gene were successfully amplified employing published primers. At promoter region 1 (24 bp indel at -358) of prolactin gene, the proportions of DD and ID genotypes were higher in Giriraja (0.45) and Indigenous (0.44) chicken, respectively. However, the frequency of D allele was higher in both the studied populations. The restriction enzymes, AluI and Csp6I were utilized for genotyping polymorphisms of C -2402T and C -2161G, respectively. At C -2402T, TT and CT genotypes were predominant in Giriraja and Indigenous chicken with frequencies of 0.61 and 0.56, respectively. The frequency of T allele was higher in both the studied populations. Whereas at C-2161G, GG and CG genotypes were predominant in Giriraja (0.49) and Indigenous (0.65) chicken, respectively. In both the studied populations the frequency of G allele was higher. Further, three SNPs viz., C>T transition at 161 bp, T>C transition at 371 bp and C>G transversion at 402 bp were detected at promoter region 2 (C-2402T and C-2161G) in T/ G allele compared to C allele.ThesisItem Open Access SPERM DNA FRAGMENTATION AND SIRE CONCEPTION RATE IN HOLSTEIN FRIESIAN BULLS(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) SANTHOSH, G. R.; NAVEEN KUMAR, G. S.The present study was undertaken to know the association of sire conception rate with bull age, sperm phenotypic characters, sperm DNA fragmentation (SDF), protamine polymorphism and PRM1:PRM2 expression ratio in selected HF bulls maintained at State Semen Collection Center, Bengaluru, Karnataka. The overall mean sire conception rate in the study was 50.61 ± 1.17%. Group G-I (Bulls aged < 3 years) had significantly lower sire conception rate (45.0 ± 0.01%), than G-II (3-5.5 years) and G-III (> 5.5 years) with sire conception rates of 50.61 ± 1.17% and 53.0 ± 0.017% respectively. The Single Nucleotide Polymorphism (SNP) analysis of Protamine1 gene revealed presence of six different SNPs in the 5’ promoter region and no SNPs in the exonic and intronic region of Protamine 1 gene. The SNP in TATA box TT > TG (TATATAA), 30 bp upstream from exon 1 was found to be related to the low expression of Protamine1 and hence a lower sire conception rate. No differences in semen and sperm phenotypic traits were observed among bulls of different age group. The average PRM1 to PRM2 ratio and DNA Fragmentation Index% (DFI) observed in present study were 1:0.06 and 8.88 ± 0.22% respectively. Significant lower expression levels of PRM1 was observed in younger (GI) bulls. DFI% was higher in younger aged bulls (9.2 ± 0.39) than aged bulls (8.6 ± 0.36) but not significantly different. DFI% was negatively correlated with PRM1 (r = -0.660) and PRM2 (r = -0.518) expression levels. Sire conception rate was negatively correlated with DFI% also and positively correlated with PRM1 expression (0.564) and Protamine ratio (0.581). However, PRM2 was found to be not significantly correlated with sire conception rate. Present study revealed that lower protamination is a probable reason for lower sire conception rate in younger bulls.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.ThesisItem Open Access MOLECULAR CHARACTERIZATION OF MBL1 GENE AND ITS ASSOCIATION WITH SOMATIC CELL COUNT IN DEONI AND CROSSBRED CATTLE(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2019-09) KHAMGAL PRASHANT RAMCHANDRA; YATHISH, H. M.)The present investigation was undertaken to explore the nucleotide variabilities in coding regions of MBL1 gene using PCR-RFLP and to associate them with somatic cell count (SCC) in 64 Deoni and 50 HF crossbred cattle. In Deoni cattle, only MBL1_E5/StyI analysis revealed AA and AB genotypes, and remaining MBL1_E1/HinfI, MBL1_E2/MslI, MBL1_E3/MmeI and MBL1_E4/BcoDI analysis revealed monomorphism. The frequency of genotypes was varying from 0.17 to 1.0 and that of ‘A’ and ‘B’ alleles was varying from 0.41 to 1.0 in Deoni cattle. In HF Crossbred cattle, MBL1_E2/MslI, analysis revealed AA and AB genotypes, and remaining MBL1_E1/HinfI, MBL1_E3/MmeI, MBL1_E4/BcoDI and MBL1_E5/StyI analysis revealed monomorphism. The frequency of genotypes was varying from 0.28 to 1.00 and that of ‘A’ and ‘B’ alleles was varying from 0.36 to1.00 in HF crossbred cattle. Sequence analysis, when compared to Bos taurus sequence, has revealed three SNPs viz. T244C, G392A and G437A and two SNPs viz. G209A and T244C in ‘A’ allele of MBL1_E2/MslI in Deoni cattle and HF Crossbred cattle, respectively. χ 2 -square analysis revealeThesisItem Open Access GENETIC STUDIES ON LOCAL GOATS IN MANDYA DISTRICT OF KARNATAKA(KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2016-05) SIDDALINGA MURTHY, H. K.; (M.R. JAYASHANKAR; K. ANIL KUMAR); C.S. NAGARAJA; H.N. NARASIMAHA MURTHY; K. SATHYANARAYAN; R. BHASKARANA study was undertaken on local goats in Mandya district of Karnataka for various morphological characters, production and reproduction parameters, heritability of body weight at different ages and to develop prediction equations for adult body weight. Characteristic features of the goats were predominantly black colour of coat, eyelid, muzzle and hooves, leafy structured and pendulous ears, straight forehead, absence of beard and wattles and medium sized tail. Among horned goats 76.20 per cent had straight horns with upward backward orientation. Overall least squares means for body weight at three, six, nine, twelve month and adult goats were 9.41±0.12, 15.44±0.09, 19.46±0.09, 21.87±0.09 and 27.20±0.05 kg, respectively. Males were significantly (P≤0.01) heavier than females in all age groups. Single born kids were heavier than twin and triplet born kids. Effects of sex, type of birth and location/ centre on all body measurements were significant. Overall mean age at first conception, age at first kidding, kidding interval and service period in Mandya local goats were 398.94±3.31, 550.07±3.31, 276.00±2.17 and 125.47±0.93 days, respectively, and were significantly affected by year, season and type of birth. Heritability estimates of body weight were 0.36±0.03, 0.29±0.02, 0.27±0.04 and 0.24±0.02 at three, six, nine and twelve months, respectively. Body weight was positively and highly significantly correlated with all the four body measurements viz., height at withers (0.80), body length (0.96), chest girth (0.94) and paunch girth (0.94). Multiple regression analysis indicated that Height at Withers (X1) in combination with Body Length (X2) and Chest Girth (X3) would significantly improve accuracy of predictions.ThesisItem Open Access Study on Association of Growth Hormone Gene Polymorphism with Milk Production in Cattle(Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar, 27-12-12) Punya, J.; Jayashankar, M.R.; Girish Kumar, V.; Shrikrishna Isloor; Nagaraja, C.S.The genetic variation at the growth hormone gene in a HF cross bred population and to study the possible association of the identified polymorphism with first lactation milk yield. Two fragments of bovine growth hormone gene covering part of intron 3 and exon 5 were PCR amplified from 50 HF cross bred cattle and subjected to RFLP with MspI and AluI enzymes respectively. PCR-RFLP of exon 5 of growth hormone gene with AluI yielded monomorphic pattern of 223, 171 and 52 bp fragments. The sequencing of PCR product confirmed the pattern as heterozygous (LV). No homozygotes were observed in the population. The PCR-RFLP of intron 3 region of growth hormone gene with MspI yielded three genotypes. The frequency of genotypes Msp (+/+), Msp (+/-) and Msp (-/-) were 0.28, 0.12 and 0.60 respectively. The population was not in Hardy Weinberg Equilibrium with respect to Msp polymorphism. The average first lactation milk yield of HF cross bred cows with genotypes Msp (+/+), Msp (+/-) and Msp (-/-) were 2493 kgs, 2473 kgs and 2558 kgs respectively and were not significantly different indicating that the SNP had no influence on the first lactation milk yield in the population studied.
