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Cytochrome b is the central catalytic subunit of the quinol : cytochrome c oxidoreductase of complex III of the mitochondrial oxidative phosphorylation system and is essential to the viability of most eukaryotic cells. Partial cytochrome b gene sequences of 14 species representing mammals, birds, reptiles and amphibians are presented here including some species typical for Poland. For the analysed species a comparative analysis of the natural variation in the gene was performed. This infor­mation has been used to discuss some aspects of gene sequence — protein function relationships. Review of relevant literature indicates that similar comparisons have been made only for basic mammalian species. Moreover, there is little information about the Polish-specific species. We observed that there is a strong non-random dis­tribution of nucleotides in the cytochrome b sequence in all tested species with the highest differences at the third codon position. This is also the codon position of the strongest compositional bias. Some tested species, representing distant systematic groups, showed unique base composition differing from the others. The quail, frog, python and elk prefer C over A in the light DNA strand. Species belonging to the ar- tiodactyls stand out from the remaining ones and contain fewer pyrimidines. The ob­served overall rate of amino acid identity is about 61%. The region covering Qo cen­ter as well as histidines 82 and 96 (heme ligands) are totally conserved in all tested species. Additionally, the applied method and the sequences can also be used for di­agnostic species identification by veterinary and conservation agencies.
Recently we have shown that in vitro binding of the proximal part of the human tyro­sine hydroxylase gene to the nuclear matrix is correlated with its transcriptional activ­ity. The strongest binding potential was predicted by computing for the first intron se­quence (Lenartowski & Goc, 2002, Neurosci Lett.; 330: 151-154). In this study a 16 kb fragment of the bovine genomic DNA containing the tyrosine hydroxylase gene was investigated for its affinity to the nuclear matrix. Only a 950 bp fragment encod­ing the distal part of the first intron, second exon and a few nucleotides of the second intron bound to the nuclear matrix. The binding was independent of the tissue-specific tyrosine hydroxylase gene activation. The fragment was subcloned and sequenced. Computer search pointed to one potential intronic matrix attachment region with two AP1-like sites embedded in the sequence. We conclude that even if the position of the matrix binding region is conserved among the tyrosine hydroxylase genes in mam­mals, its tissue specificity and/or function is not preserved or is achieved by different mechanisms.
The purpose of the study was to elaborate a molecular procedure to identify the DUMPS mutation which causes hereditary disorder (deficiency of uridine monophosphate synthase) in cattle. The degree of this disorder was determined in a gene pool of a Polish bredcattle population which consisted of 1038 animals: 781 artifical insemination bulls from all centers in Poland and 257 sire dams from the two largest breeding centers of Holstein-Friesian cattle: Osięciny and Osowa Sień. The diagnosis was performed using the PCR-RFLP method and technique based on automated solid-phase sequencing of PCR products. In the examined material no DUMPS carrier was found. This indicates that the population of cattle reared in Poland is free from this economically significant disorder. The results obtained indicate a high effectiveness of monitoring of point mutation DUMPS by the PCR-RFLP method. Moreover, it is worth noting the wide possibilities of examining other biologically significant genome regions using the solid-phase sequencing method.
Analysis of microsatellite instability (MI) and loss of heterozygosity (LOH) is rec­ommended for screening patients with sporadic and hereditary malignancies. This study shows an application of a fluorescent hexaplex PCR system for microsatellite typing on A.L.F. DNA Sequencer (Pharmacia Biotech). This technique detects changes in microsatellites providing a time-efficient, reliable and accurate method for MI and LOH analyses. The Fragment Manager software was used for automated size calculation and quantitation of DNA fragments, enabling rapid and precise mea­surement of allelic ratios. We examined 70 breast cancer and 70 control DNA speci­mens, classified all the patterns of microsatellite alterations, and set up MI and LOH assessment criteria for the automated multiplex fluorescent method.
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