Pituitary 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 Simmen...tal, 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.
The 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.
ITS (internal tanscribed spacer) profiles of the aDNA (ancient DNA) of seed remains extracted from an extinct sample recovered from the 15th century (Budapest, Hungary) were compared to 31 modern melon cultivars and landraces. An aseptic incubation followed by ITS analysis was used to exclude the exogenously and endogenously contaminated (Asper...gillus) medieval seeds and to detect SNPs in ITS1-5.8S-ITS2 region of rDNA (ribosomal DNA). SNPs were observed at the 94–95 bp (GC to either RC, RS or AG) of ITS1; and at 414 bp (A-to-T substitution), 470 bp (T to Y or C), 610 bp (A to R or G) of ITS2. The results facilitate the final aim of molecular and morphological reconstructions of ancient melon tpyes.
Seed 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 b...y 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.
In 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 p...rimers 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.
In 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 ex...plored.