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Effect of G2548A polymorphism in the leptin gene on the BMI level in human population
5-10Views:162The polymorphism in leptin (LEP 2548A) seems to influence obesity among others genes. The aim of this study is to investigate the effect of the G2548A polymorphism on body mass index. We included 79 people from Slovakia with some genetic relatedness and used barrels kit to isolate the genomic DNA from an adenoblast swab- from the salivary. PCR products were amplified by pursued polymorphisms and G2548A, we restriction-analyzed them and then we identified the specific fragments describing the presence of chosen SNP polymorphism by the agarose electrophoresis, to analyze SNP polymorphism by PCR-RFLP method.
The LEP gene had increased frequency of G allele (0.5506). The most common genotype occurring in the gene LEP was heterozygous genotype (AG) and the least frequent genotype in LEP was AA (0.1899). Taking the age into account the BMI is higher if the G allele occurs in the LEP gene. Moreover, if the G allele genotype was situated in dominant form, then the highest average BMI was present.
According to the results we can assume that the AA genotype (LEP) has a protective effect on the prevalence of obesity compared to the other genotypes.
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Association analysis of TNNI1/XbaI polimorphism on carcass quality in hybrid pigs
59-62Views:199The contractile protein, which is encoded by troponin I 1 (TNNI1) gene, is located on the thin filaments of slow fibres in striated muscle. TNNI1 protein is a part of the troponin complex which plays an important role in regulation of muscle contraction by preventing actin-myosin interaction in absence of calcium. According to biological role, this gene can be potential marker for meat production related traits. The aim of this study is to define whether the previously reported gene polymorphism (EU743939:g.5174T>C) is connected with the slaughter traits measured in a standard slaughterhouse of the examined four-line European hybrid. The study included data from 404 gilts and barrows from 2 different samples. The polymorphism was detected using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) method with XbaI restriction enzyme. In this study the allele frequencies were found as follows: C: 0.84 and 0.808; T: 0.16 and 0.192. Based on result of the present study no significant impact of polymorphisms on production parameters was found.
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Associations analysis of production traits with leptin gene T3469C polymorphisms in pig
39-43Views:201The aim of this study was to define the connection between the leptin (LEP) gene T3469C polymorphism and its potential association with production traits in improved hybrid pigs. The study included data from 397 gilts and barrows from 2 different sample. The polymorphism was identified by using the polymerase chain reaction-restriction fragment-length polymorphism (PCR–RFLP) method with HinfI restriction enzyme. Two alleles of LEP gene were identified: T (0.93) an C (0.07). The analysis of values of production traits, depending on LEP genotype did not reveal significant (P≤0.05) differences. In the examination the loin diameter (between the 2nd and 3rd ribs), the live weight at slaughter and the averege daily gain during fattening were higher at pigs with C allele than pigs with TT genotipe. Accordingt to our data the effect of C allele was favourable in this population, because these animals had bigger bodyweight without valuable change of lean meat percent.
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Brief overview of the polymorphism analysis of genes affecting pig prolificacy (LEP, PRLP, ESR BF, EGF, FSH-β, H2A.Z)
5-10Views:187Researches are being performed around the world to increase swine prolificacy by using marker-assisted selection (MAS). The present study processes researches of polymorphism examinations on 7 genes. The result of the experiments showed that the leptin gene (LEP) prolactin receptor gene (PRLP), estrogen receptor gene (ESR), properdin B (BF) epidermal growth factor (EGF), follicle-stimulating beta gene (FSH-ß) and Z member of the H2A histon family gene (H2A.Z) and their alleles have a positive effect on reproductive characteristics of different swine breeds. In addition to this, leptin gene (LEP) influences the build, meat production and growth of body fat. Further studies are concerned with the polymorphism of an increasing number of genes, which enables a faster genetic development of swine breeding.
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Polymorphism of the bovine GH and LEP genes in a population of Slovak spotted bulls
19-23Views:107The aim of this study was the detection of polymorphism in the bovine growth hormone and leptin genes using the PCR-RFLP method. A
polymorphic site of the growth hormone gene (Alul loci) that results in amino acid change at position 127 of the protein chain (leucine, L to
valine, V) has been linked to differences in circulating metabolites, metabolic hormones and to milk yield. The polymorphism in bovine leptin
gene is situated in the intron between two exons, which results in an amino acid change at position 2059 of the protein chain (cytosine, C to
thymine, T). The polymorphisms were studied in a group of 58 bulls of the Slovak spotted breed. A strategy employing PCR was used to amplify 428 bp (GH gene) and 422 bp (LEP gene) products from blood samples. Digestion of PCR products with restriction enzymes AluI and Sau3AI revealed alleles: L and V; A and B for GH gene and LEP gene, respectively. The growth hormone gene is a candidate gene for body weight gain in cattle, since it plays a fundamental role in growth regulation. Leptin plays an important role in the regulation of feed intake, energy metabolism, growth and reproduction of cattle; therefore, animals with higher leptin gene expression will probably have lower daily weight gain than others with similar forage offer and nutritional condition and will also likely have longer calving intervals. -
The results of the investigation of the β-casein gene polymorphism in sheep breed – Review
75-80Views:240β-casein is the most abundant protein fraction in sheep milk, and has at least six different alleles (A, B, C, G, X, Y). The alleles of the β-casein gene may influence on the quality and quantity of milk. Knowing the gene polymorphism has an important role in the process of milk production. The properties of milk could be positively influenced by themolecular genetic methods.
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Investigation of the β-casein (CSN2) gene polymorphism in goat breed – Review
69-73Views:308Milk plays an important role in the human nutrition as an important protein and energy source. Β-casein is the most abundant casein fraction in the goat milk. The β-casein gene has at least 9 polymorphism in goat breed (A, A1, B, C, C1, D, E, 0 and 0’). The most important alleles are the zero (0) and zero’ (0’) alleles, because they decrease the quantity of β-casein protein in milk. The lower quantity of the β-casein changes the properties of the coagulation and the allergenin milk. This review is about the β-casein gene polymorphism in goat breed.
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Search for polymorphism in exon 5 of cattle pituitary adenylate cyclase activating polypeptide gene
17-20Views:199Pituitary adenylate cyclase-activating polypeptide is a neuropeptide expressed primarily in the hypothalamus and in other tissues, which divergent and extensive physiological functions are proved. Large number of SNPs in PACAP are published involved SNP that is associated with phenotypic trait of cattle as well. Hungarian Grey, Hungarian Simmental, Holstein, Charolais and Angus cattle breeds were involved in this study to search for polymorphism in exon 5 1–391 bp and G/A transition. Our results of PCR RFLP and PCR SSCP did not prove the occurrence neither G/A transition, nor any other SNP in cattle breeds involved.
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Genetic polymorphism of candidate genes in pig meat production
37-40Views:105H-FABP, LEPR and MC5R genes were suggested as candidate genes for fat content in pig meat. The aim of this study was to detect genetic variation in the porcine H-FABP, LEPR and MC5R genes by PCR-RFLP method in a group of pigs. Genotyping of pigs was done by PCRRFLP methods. We identified three genotypes in the set of pigs, HH (0.504), Hh (0.412) and hh (0.084) for H-FABP (HinfI). Allele H showed higher frequency than allele h (0.710 vs. 0.290). Three genotypes were identified for the H-FABP (HaeIII) gene (DD - 0.194, Dd - 0.494, dd - 0.312). The allele D (0.441) showed slightly lower frequency than allele d (0.559). All three genotypes were identified for LEPR (HpaII) in the group of pigs (AA – 0.137, AB - 0.314, BB – 0.549). Higher frequency of LEPR gene was confirmed for allele B (0.706), as compared with allele A (0.294). We identified two genotypes for MC5R (BsaHI) in the group of pigs (AA - 0.348 and AG - 0.652), genotype GG was not found. As conforms with genotype structure, we recognize a higher frequency of allele A (0.674) as compared with allele G (0.326).
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Sequence stability at SSR, ISSR and mtDNA loci of common millet (Panicum miliaceum) from the middle ages
10-19Views:108Seed remains of medieval millet, recovered from a 15th century layer (King’s Palace, Budapest, Hungary), showed reddish yellow grain color after rehydrating on tissue culture medium that was close to grain color of modern cultivar Omszkoje. aDNA of medieval c. millet was extracted successfully, analyzed and compared to modern common millets by ISSR, SSR, CAPS and mtDNA. Analyses of fragments and sequences revealed
polymorphism at seven ISSR loci (22 alleles) and at the 5S-18S rDNA locus of mtDNA. CAPS analysis of the 5S-18S rDNA fragment revealed no SNPs in the restriction sites of six endonucleases TaqI, BsuRI, HinfI, MboI, AluI and RsaI. Sequence alignments of the restriction fragments RsaI also revealed
consensus sequence in the medieval sample compared to a modern variety. Morphological characterization of twenty common millet (Panicum miliaceum L., 2n=4×=36) cultivars and landraces revealed four distinct clusters which were apparently consistent with the grain colors of black, black and brown, red, yellow, and white. In the comparative AFLP, SSR and mtDNA analysis modern millet cv. ‘Topáz’ was used. AFLP analysis revealed that extensive DNA degradation had occurred in the 4th CENT. ancient millet resulting in only 2 (1.2%) AFLP fragments (98.8% degradation),
compared to the 15th CENT. medieval millet with 158 (40%) fragments (60% degradation) and modern millet cv. ‘Topáz’ with 264 fragments (100%). Eight AFLP fragments were sequenced after reamplification and cloning. Microsatellite (SSR) analysis at the nuclear gln4, sh1, rps28 and rps15 loci of the medieval DNA revealed one SNP (single nucleotide polymorphism) at the 29th position (A to G) of rps28 locus compared to modern millet.
Mitochondrial (mtDNA) fragment (MboI) amplified at the 5S-18S-rDNA locus in the medieval millet showed no molecular changes compared to modern millet. The results underline the significance of survived aDNA extraction and analysis of excavated seeds for comparative analysis and molecular reconstruction of ancient and extinct plant genotypes. An attempted phenotype reconstruction indicated that medieval common millet showed the closest morphological similarity to modern millet cultivar Omszkoje. -
Influence of H-FABP gene polymorphisms on slaughter value of hybrid pigs
55-60Views:243The H-FABP gene was defined as a potential candidate gene influencing the fat deposition traits, primarily the intramuscular fat content. The aim of this study is to define whether the previously reported gene mutations are connected with the slaughter traits measured in a standard slaughterhouse. The study included data from 405 gilts and barrows from 2 different samples. The two chosen mutation (HFABP1: c. 103 T>C, HFABP2: c. 1970 T>C) were detected in one reaction with PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Lenght Polymorphism) method with HinfI restrictoin enzyme. The allel frequencies are as follows: 103T(H)=0.75; 103C(h)=0.25, 1970T=0.32; 1970C=0.68. A HFABP1 mutation has significant effect on backfat thickness and lean meat % at stable 1 (sample 1), but there were no effect at stable 2 (sample 2). The analysis of values of production traits, depending on HFABP2 genotype did not reveal significant differences. Based on this study we can’t get a clear conclusion on the impact of polymorphisms on production parameters. In the examined flock the allele frequency of mutation in 5 'UTR is identical to the literature data, i. e. the more favorable variant regarding the intramuscular fat content is predominant in the population.
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Lack of polymorphism of the agouti signaling protein (ASIP) gene among four different brown hare (Lepus europaeus Pallas 1778) populations
81-85Views:164The brown hare (Lepus europaeus Pallas 1778) is a common palearctic and a popular game species therefore it has been an obvious subject for population genetic studies since the second part of the 20th century. Among the several mitochondrial DNA studies some have been carried out concerning nuclear genes as well. The agouti signaling protein gene (ASIP) is involved in regulating the synthesis of eumelanin and pheomelanin in melanocytes of mammals. Though many studies focused on it in relation with several mammalian species, minimal information is available on this topic concerning the brown hare.
Here we present a short communication concerning the agouti signaling protein (ASIP) gene in four different country’s L. europaeus populations, namely Lithuania, Hungary, Serbia and Georgia. N=45 tissue samples have been investigated from overall 17 sampling sites of the different countries. There has not been found any polymorphism among the sequences. In an alignment with other Leporid species’ partial ASIP sequences downloaded from ENA we have found that based on a 178 base pairs long DNA sequence the haplotype of our samples contains three other Lepus species as well. This is concordant with the findings of a previous study focusing predominantly on the European rabbit (Orycto lagus cuniculus Linnaeus 1758) and the several mutations of its ASIP gene.
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Investigation of the heat shock gene Hsp70 polymorphism in different sheep genotypes
41-45Views:184Nowadays the climate change has an increasing effect on the animals. The warming climate brings up several problems on the area of the animal husbandry, which ones are really important. From the first time the living beings have defensive mechanisms against the heat shock. In current examination we use– from Hungary and from other countries collected –samples of sheep breeds, which are living on different climate. Our fundamental assumption was, that the animals living on other climate adapted to the changes of the environment and there are differeces in their genetic background. These fixed mutations we are looking for in the HSP70 heat shock gene, but we haven’t found any polymorphism yet. We are going to involve further breeds and more individuals in the investigations.
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Single nucleotide polymorphism analysis in meat-production related genes in broiler chickens
79-82Views:184In broiler chickens, the intensive selection for growth rate, feed efficiency, body composition (breast muscle weight) traits in the last decades was successful. To improve economically important characteristics, it is possible to use molecular markers associated with meat production traits. The aim of this study was to examine genotype polymorphisms in ROSS 308 broilers for thyroid hormone responsive Spot14α, insulinlike growth factor 1 (IGF1), IGF-binding protein 2 (IGFBP2), somatostatin (SST) and prolactin (PRL) genes. A further goal of this investigation was to study the relationship between the polymorphisms and phenotypic characteristics.
In the investigated broiler population, the frequency for CC homozygous genotype was 0.77 in Spot14α (AY568628), AA homozygous genotype was 0.80 in IGF1 (M74176), GG homozygous genotype was 0.85 in IGFBP2 (U15086), DD homozygous genotype was 0.60 in PRL (FJ663023 or FJ434669). Only the AA homozygous genotype was found in SST (X60191). Chickens with AC genotype in Spot14α, and with GG genotype in IGFBP2 had higher body weight (BW) and carcass weight (CW), compared to CC and GT genotypes. However, the differences were not significant (P>0.05). There was significant association (P<0.05) between PRL genotypes and body and carcass weight, where chicken with homozygous DD surpassed individuals with homozygous II genotypes.
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Detection of DNA mutations by PCR-TTGE method
21-25Views:212In our study PCR-temporal temperature gelelectrophoresis (TTGE) and MeltINGENY bioinformatic program were used to analyse the mutations in the genes of melanocortin-1 receptor (MC1R) and pituitary adenylate-cyclase activating polypeptide (PACAP) in cattle. Amplification of target DNA by PCR was performed with GC-clamp primers and non-GC-clamp primers in simplex PCR reactions. The fragments were separated by denaturing polyacrylamide gelelectrophoresis (denaturing agents: high temperature, urea) after PCR reactions.MC1R homozygous individuals were used for the reaction.
We concluded that MeltINGENY program makes the decision and detection system easier, and more simple as the melting profile of target sequence is determined by the software. In case of MC1R gene, PCR-TTGE method is appropriate for SNP detection, however PACAP gene polymorphism can not be identified by the method, because PACAP mutations are not included in melting domains, therefore PCR-TTGE cannot detect them.
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Investigation of genetic diversity in irradiated maize lines and its relation to hybrid performance
20-26Views:227Knowledge of genetic diversity among available parental lines is fundamental for successful hybrid maize breeding. The aims of this study were to estimate (1) genetic similarity (GS) and genetic distance (GD) (based on Jaccard index) in four maize inbreed lines; (2) to classify the lines according to their GD and GS; (3) to determine hybrid performance based on GD and heterosis for yield ability in 4x4 full diallel system. We used morphological description and AFLP (amplified fragment length polymorphisms) for estimation genetic polymorphism in four maize inbred lines. We estimated the applicability of genetic similarity in SC and reciproc hybrids for prediction of their performance.
Three primer combinations were used to obtain AFLP markers, producing 207 bands, 70 of whit were polimorphic. The dendogram based on genetic similarities (GS) and genetic distance (GD) and morphological description separated four inbred lines into well-defined groups. Morphological description just with AFLP analysis showed reliable results. In view of genetic distance, the UDL 1 line and their linear and reciprocal crosses showed significant heterosis effect, which was confirmed by heterosis calculation based on grain yield. -
Examination of the interval between litters (IBL) of different genotype HLW sows using survival analysis
13-17Views:243In this study our aim was to find out if there is a difference between the genotypes determined for the previously identified mutations of seven genes of the Hungarian Large White in terms of the time spent in production. We identified the previously determined alleles of the seven genes (BF, EGF, ESR, FSHβ, H2AFZ, LEP, PRLR) related to proliferation that were and performed the survival analysis between breeds indicating the risk of culling and the time spent in production on the given farm. Based on the results of survival analysis by Log-rank test, Breslow (Generalized Wilcoxon) and Tarone-Ware test we concluded that they indicated a significant difference in case of the genes BF (Breslow and Tarone-Ware tests) the EGF (Log-rank and Tarone-Ware tests) and ESR (Log rank test) based on which the curves of the survival of the certain genes varied form one another significantly.
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Introduction of DNA-based Methods to Agriculture Through Molecular Taxonomic Examination of Poa Species
139-142Views:114biological methods, one among them is AFLP that is well applicable for taxonomic research. Bluegrass species, that are important components of meadow associations, thus their thorough knowledge is necessary in maintaining biodiversity, were examined with bringing this method to perfection.
Taxonomic relationship of the members of Poa pratensis aggregation is a controversial issue. Present study aimes to identify the members of this group, with a developed AFLP method through molecular taxonomic examination of Poa species in meadows nearby Debrecen, revealing their genetical distances. Species of the aggregation show a great genetic variability, but their genetic proximity approves the use of the term aggregation. Results established wider geographical investigation of three species of the aggregation. The distinctness of the species based on their morphological features was confirmed according to their genetical basis as well. The method overall turned out to be appropriate for the taxonomic research of bluegrass species like Poa pratensis and its aggregation. -
Molecular Biological Approach of Crocus sativus L. and its Allies
112-115Views:168The hay saffron (Crocus sativus L.) is a sterile triploid plant, known in human culture only, with no fertile seeds produced. The origin of saffron is still a mist, however it is assumed to be an autopoliploid mutant or a hybrid. The recent classification and most of the former taxonomic publications define C. sativus to be derived from C. cartwrightianus, a wild species. Because of the sterility of hay saffron it seemed to be reasonable to apply molecular biological methods to complete classical taxonomic studies in examining its relations. The DNA polymorphism based AFLP method has confirmed the close relationship between these species.
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Application of molecular genetic methods for species Equus caballus – Literature review
21-28Views:175In this study our aim was to provide a comprihensive overview of the most commonly used methods in molecular genetic studies related to Equus caballus. Thus we are dealing with the D-loop region of mitochondrial DNA, with microsatellites and also with single nucleotid polimorphism as SNP. The advantages and drawbacks of each method were also explored.
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Application of AFLP-Method in Plant Sample Identification
207-213Views:82One possible method for the determination of DNA-polymorphism is the PCR-based AFLP (Amplified Fragment Length Polymorphism). This method had been succesfully introduced to the Department of Botany at University of Debrecen in 2000-2001 with the examination of hay saffron (Crocus sativus L.) and its allies. Hay saffron is grown as a spice for some thousand years producing the most expensive spice in the world. This plant is sterile, triploid reproduces only vegetatively with no fertile seeds. However its origin is unknown it exists only in cultivation and it is a mutated variety of another species or an artificial or natural hybrid. Usual methods for the systematic examination are restricted hence it seemed to be reasonable to apply molecular biological methods in its case. Results of this work include the introduction and many fold application of the method beside ensuring the consequences of science literature with determining the C. cartwrightianus to give the most similar genetical pattern to C. sativus.