The aim of this thesis was to analyze the polymorphism in position -371bp (related to ATG start codone) of MSTN gene, SNP in position 1984bp of MYF5 gene and influence of these polymorphisms on tenderness, water holding capacity, pH and color of meat. Samples (241) of bull meat of Czech pied cattle were used for analysis. PCR-RFLP method was applied to genotype MSTN and MYF5 genes. For polymorphisms detection was used restriction endonucleases DraI (for MSTN gene) and TaqI (for MYF5 gene). From established genotypes were computed their frequencies and alleles frequencies. The frequencies of genotypes in MSTN gene were AA=0,729, AB=0,258, BB=0,013 and alleles frequencies were A=0,858, B=0,142. The frequencies of genotypes in MYF5 gene were AA=0,181, AB=0,542, BB=0,278 and alleles frequencies were A=0,452 a B=0,548. Using statistical analysis, the influence of genotypes of MYF5 gene on the water holding capacity, pH and the color of meat was determined.

Congenital hypotrichosis in cattle is commonly referred to as "rat-tail" syndrome and is characterized by a dilution of black coat color and morphological changes to the hair shaft and tail switch. Two loci are involved in the inheritance of the rat-tail phenotype, the "extension locus" (MC1R) and an unknown locus. In order to express the rat-tail phenotype the animal must inherit at least one black allele at MC1R and be heterozygous at the unknown locus. The rat-tail locus was previously mapped to an 8.7 cM region of Bos Taurus autosome (BTA) 5. Pmel17 is known to be involved in the expression of pigmentation and maps to the same region of BTA5 as the rat-tail locus. Cattle from a population segregating for the rat-tail syndrome were sequenced at Pmel17 in order to identify putative causative mutations. Two mutations were detected, a three base pair (bp) deletion in exon 1 at codon 18 removing a leucine residue, and a single nucleotide polymorphism (SNP) at codon 612 resulting in an amino acid substitution (A?E). The 3-bp deletion in exon 1 of Pmel17 is in 100% concordance with the rat-tail phenotype in this research population and may be causative of the rat-tail phenotype.

No description available.

No description available.

No description available.

La transition florale est un évènement clef dans la vie d’une plante. Chez le fraisier, la compréhension des mécanismes génétiques de cette transition est un enjeu majeur pour mieux contrôler la production de fruits. La transition florale peut être étudiée à travers la remontée florale, qui est la capacité d’une plante à fleurir tout au long de la période végétative. Le fraisier cultivé octoploïde, F. x ananassa, comme le fraisier diploïde, F. vesca, présentent des génotypes remontants capables de fleurir en continu. L’objectif de cette thèse est de comprendre le déterminisme génétique et moléculaire de la remontée florale chez Fragaria. Ce travail a montré que chez les fraisiers diploïde et octoploïde, le caractère ‘remontée florale’ est contrôlé par deux verrous génétiques différents localisés à des positions non orthologues. Chez le fraisier diploïde, le gène FvKSN responsable de la remontée florale a été identifié et code pour un homologue du répresseur floral TFL1. Chez les génotypes remontants, ce gène présente une délétion dans la partie codante conduisant à une protéine non fonctionnelle, incapable de réprimer la floraison. Chez le fraisier octoploïde, le QTL majeur détecté contrôlant la remontée florale est lié à la production de stolons de manière antagoniste, suggérant l’existence d’une région génomique où s'exerce une compétition entre multiplication végétative et la reproduction sexuée. Cette région génomique comprend plusieurs gènes candidats intéressants dont FT, activateur de la floraison.Une hypothèse suggérée par ce travail est que chez le fraisier, l’alternance entre phase végétative et phase reproductive est liée à l’équilibre entre les gènes FvKSN, homologue de TFL1, et FvFT, homologue de FT. La remontée florale serait la conséquence d’une modification de cet équilibre entre ces deux gènes en faveur du développement reproductif. / The floral transition is a key event in plant life. In strawberry, understanding the genetic mechanisms of floral transition is a major issue for better control of fruit production. This transition is studied through the continuous flowering, which is the ability to flower throughout the growing season. Both, the octoploid cultivated strawberry, F. x ananassa, as the woody diploid strawberry, F. vesca, displayed continuous flowering genotypes. The objective of this work is to decipher the genetic and molecular mechanism of the continuous flowering in Fragaria.This work has shown that in diploid and octoploid strawberry the continuous flowering is controlled by two different genetic 'keys' located at non-orthologous position. In diploid strawberry, the gene FvKSN responsible of continuous flowering was identified and encodes a homologous to the TFL1 floral repressor. In the continuous flowering genotypes, this gene has a deletion in the coding region leading to a nonfunctional protein unable to repress flowering. In the octoploid strawberry, the major QTL controlling both the recurrent flowering and the runner production was identified. These traits were antagonist, which suggests competition between vegetative propagation and sexual reproduction in this region. This genomic region contains several interesting candidate genes whose FT, an activator of flowering.A hypothesis could be proposed. In strawberry, the switch between vegetative and reproductive phase is linked to balance between two genes, FvKSN, homologous to TFL1 and FvFT homologous to FT. Continuous flowering would be the consequence of balance modification between this two genes to the benefit of floral development.

Lowering LDL-cholesterol with statin therapy produces substantial reductions in cardiovascular events, and larger cholesterol reductions may produce larger benefits. Rarely, myopathy occurs with statins, especially at higher doses and in combination with certain medications. Similarly strong associations might exist between myopathy with high-dose statin regimens and genetic variants, especially those affecting blood statin levels. This study aimed to find genetic variants associated with statin-induced myopathy. A feasibility study was completed to assess whether plausible effect sizes of 5 to10-fold higher risks per genetic variants could be detected among 50-100 cases with statin-induced myopathy and to consider the best study design. A genome-wide association study was then carried out using approximately 300,000 genetic markers (and additional fine-mapping) in 85 people with definite or incipient myopathy and 90 controls, who were all taking 80mg simvastatin daily in a 12,000 participant trial of 80mg vs 20mg simvastatin daily. The cases were also compared to 2,300 additional controls who had not been exposed to intensive-dose statin therapy. Replication of the myopathy result and lipid-lowering associations were tested in a 20,000 participant trial of 40mg simvastatin daily versus placebo. The genome-wide scan yielded a single strong association (p = 4x10^-9) of myopathy with the rs4363657 single nucleotide polymorphism (SNP) located within the SLCO1B1 gene on chromosome 12. This non-coding SNP was in nearly complete linkage disequilibrium (r²=0.97) with the non-synonymous rs4149056 SNP. The population prevalence of the rs4149056 C allele was 15%, and the odds ratio for myopathy was 4.5 (95% confidence interval 2.6 to 7.7) for each copy of the C allele and 16.9 (4.7 to 61.1) for CC vs TT homozygotes. Over 60% of these myopathy cases could be attributed to the C variant. The SLCO1B1 gene encodes the organic anion transport polypeptide OATP1B1, which has been shown to regulate hepatic uptake of statins. In literature reports, rs4149056 reduced statin transport and was associated with 37% (31% to 44%) higher systemic statin acid levels per C allele. The association of rs4149056 with myopathy was replicated in the trial of 40mg simvastatin daily, which also showed that it was associated with the cholesterol-lowering effects of simvastatin. No SNPs in any other region were clearly associated with myopathy (although comparison of the myopathy cases with the 2,300 controls identified a region of chromosome 1p12 that warrants further study). This study identified common variants in SLCO1B1 that influence the risks of statin-induced myopathy substantially. Genotyping these variants may be useful for tailoring both the statin dose and safety monitoring. More generally, such studies of the genetic determinants of serious adverse reactions with other drug classes may help to improve the balance between treatment efficacy and safety.

Dentre o setor agropecuário, a avicultura é a que mais tem demonstrado índices de evolução nos últimos anos. Esses avanços são obtidos principalmente por meio da nutrição, manejo dos animais e seleção genética. A biotecnologia tem ganhado papel de destaque com o uso de marcadores moleculares como ferramenta para acrescentar informações genômicas aos processos de melhoramento convencional. Estudos anteriores em uma população F2 originada do cruzamento de frangos de corte e postura permitiram a identificação de um SNP no gene FGFBP1 (Proteína de ligação do fator de crescimento do fibroblasto 1) (g. 2014 G> A) no cromossomo 4 de Gallus gallus (GGA4). Este gene está em uma região de QTLs associado com rendimentos de coxa e sobrecoxa, peso vivo aos 35 e 41 dias de idade. O objetivo deste trabalho foi investigar um QTL previamente descrito para identificação de polimorfismos adicionais e suas associações com características de importância econômica utilizando testes de associação de um ou mais marcadores. Três genes candidatos posicionais foram identificados nesta região de QTL: KLF3(Krüeppel-like factor 3), SLIT2 (Slit homolog 2) e PPARG (Peroxisome proliferator-activated receptor gamma, coactivator 1alpha). O sequenciamento destes genes em onze (n=11) animais F1 permitiu a identificação de um polimorfismo em cada gene: g.6763 C> T (KLF3), g.187737 C> A (SLIT2) e g.76638826 -/TTTCT (PPARGC1A). Essas mutações foram genotipadas em uma população segregante F2 (n=276) e em uma linhagem pura de corte TT (n=840) da Embrapa Suínos e Aves. A frequência dos alelos para o gene KLF3 na população F2 foi de C=50% T=50% e na pura TT de C=98% T=2%, para o gene SLIT2 na população F2 foi de A=25% C=75% e na pura TT de A=30% C=70%, para o gene PPARGC1A na população F2 foi de Del=43% In=57% e na pura TT Del=33% C=67%, representando que estes polimorfismos estão segregando nas duas populações. Associações destes polimorfismos foram observadas (P<0,05) com várias características de desenvolvimento e carcaça na população F2 e na linhagem pura TT, sendo que algumas foram confirmadas nesta população como: pesos de fígado, coxas, ganhos de peso dos 35 aos 41 dias com g.6763 C> T (KLF3) e pesos das asas, cabeça, carcaça, dorso, coxas, peito, fígado e gordura abdominal com g.76638826 -/TTTCT (PPARGC1A) indicando que esta região de QTL é importante para características de produção e desempenho em frangos de corte. / Within the livestock sector, the broiler industry has showed fastest growing rates in past decades. Those advances were achieved mainly because a better understanding of the nutrition requirements, animal management and animal genetics. Biotechnology has gained a prominent role with the use of molecular markers as a tool for adding genomic information to conventional breeding processes. Previous studies using an F2 population developed from a broiler x layer cross led to the identification of a SNP on the Fibroblast growth factor binding protein 1 (FGFBP1) (g. 2014G> A) on Gallus gallus chromosome 4 (GGA4). This gene is part of a QTL associated with thigh & drumstick yields, weight gain at 35 and 41 days. This paper investigates the previously identified QTL for the identification of additional polymorphisms and their associations with important economic traits using a single and multiple markers tests. Three positional candidate genes were identified on the QTL region: KLF3 (Krüeppel-like factor 3), SLIT2 (Slit homolog 2) and PPARG (Peroxisome proliferator-activated receptor gamma, coactivator 1alpha). Sequencing of those genes was conducted in eleven (n=11) F1 animals and one polymorphism was identified in each gene g.6763 C> T (KLF3), g.187737 C> A (SLIT2) and g.76638826 -/TTTCT (PPARGC1A). These mutations were genotyped in an F2 (n=276) and a pure broiler line (n=840) from Embrapa. The frequency of the genes alleles were: KLF3 gene in F2 population C = 50% T = 50% and pure TT population C = 98% T = 2%; SLIT2 gene in F2 population A = 25% C = 75% and pure TT population A = 30% C = 70%; PPARGC1A gene in F2 population Del =43% and In = 57% and pure TT population Del = 33% C = 67% indicating that those polymorphisms are still segregating in both populations. Association was identified (P < 0,05) with several carcass and development traits in the F2 and Pure TT population, some which were confirmed like liver, drumstick weights, weight gain from 35 to 41 days with g.6763 C> T (KLF3) and wings, had, carcass, back, drumstick, breast and liver and abdominal fat weights with g.76638826 -/TTTCT (PPARGC1A) indicating that this QTL region is important for production and performance traits of broiler.

Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically useful tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of...

<p>Important candidate genes to human bone phenotypes are those involved in the regulation of hormonal action, such as the vitamin D receptor (VDR) and the estrogen receptor-α (ERα) genes and their cofactors. RIZ1 is a specific ERα cofactor proved to strongly enhance the function of the ERα. </p><p>The main focus of this thesis has been to study genetic variants in the VDR and RIZ genes and their associations to human bone phenotypes using candidate gene and functional approaches. Specifically, polymorphisms in the VDR 3’ untranslated region (UTR) and a deletion/insertion polymorphism of a proline in the RIZ gene were investigated.</p><p>The candidate gene approach was applied to large-scale population-based cohorts of pre-and post-menopausal women from Sweden and of elderly men from Sweden and Hong Kong. VDR 3’ UTR polymorphisms were associated with peak bone mass and body composition in young women. Further analysis of common VDR 3’ UTR haplotypes confirmed the association with BMD and risk of fractures in elderly men from Sweden and Hong Kong. The VDR polymorphisms were investigated for cis-acting effects, affecting allelic expression in the normal chromosomal context of human bone cells. The VDR allelic transcripts in the bone samples were unequally expressed, suggesting presence of regulatory variants in the 3’ UTR. </p><p>The polymorphism in the RIZ gene was strongly associated to BMD in pre- and postmenopausal women and in elderly men. The functional analyses included reporter constructs containing the RIZ polymorphic variants transfected in a cell line and its abilities in coactivating the ERα were examined. The variants were functionally different in coactivating the ERα-receptor complex. </p><p>To summarize, the results of this thesis show novel evidence for functional relevant polymorphisms in candidate genes to human bone phenotypes. These polymorphisms may contribute to the variation seen in BMD and risk of fractures in the population.</p>