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A single T>C nucleotide polymorphism (rs42686850) of bovine tumor necrosis factor receptor type II gene (TNF-RII) is located within a sequence with allele-specific affinity to bind E2F transcription factors, considered pivotal in the regulation of cell cycle and cell proliferation. The objective of the study was to determine the effect of this SNP and BLV infection on the TNF-RII gene expression at the mRNA and protein levels in peripheral blood mononuclear cells (PBMC). We noted that analyzed TNF-RII gene polymorphism influenced the expression of the TNF-RII gene at the mRNA level but only in BLV-positive cows. Concurrently, no statistically significant association was found between gene polymorphism and TNF-RII expression at the protein level. However, we found a significant effect of BLV infection status on the amount of TNF-RII mRNA and the percentage of PBMC expressing TNF-RII. These results show an unclear effect of considered T>C polymorphism on TNF-RII gene expression in bovine leukocytes and they suggest the involvement of BLV in modifying the TNF-RII expression in BLV-infected cows potentially implying the EBL (Enzootic Bovine Leukosis) associated pathogenesis.
Prolactin plays an important regulatory function in mammary gland development, milk secretion, and expression of milk protein genes. Hence the PRL gene is a potential quantitative trait locus and genetic marker of production traits in dairy cattle. We analysed the sequence of the PRL gene to investigate whether mutations in this sequence might be responsible for quantitative variations in milk yield and composition. Using SSCP and direct sequencing, we detected six single-nucleotide polymorphisms within a 294-bp prolactin gene fragment involving exon 4. All detected mutations were silent with respect to the amino acid sequence of the protein. PCR-RFLP genotyping of SNP 8398 R (Rsal) was used to assess allele frequencies in 186 Black-and-White cows (0.113 and 0.887 for A and G, respectively) and in 138 Jersey cows (0.706 and 0.294 for A and G, respectively). Black-and-White cows with genotype AG showed the highest milk yield, while cows with genotype GG showed the highest fat content.
In marker-assisted selection (MAS) of dairy cattle certain genes are proposed as potential candidates associated with dairy performance traits. Among different candidates, prolactin receptor gene (PRLR) seems to be promising because of its crucial role in transmitting signal from lactogenic hormones to milk protein gene promoters. In this study nine PCR fragments representing most important functional domains of PRLR were screened for polymorphism. Using SSCP method one SNP (A→C) was found in intron 9. The SNP was deposited in GenBank AY484400 and AY339393 at position of 205 nt, for Jersey and Polish Black-and-White cattle, respectively. Allele frequency was estimated in 186 Polish Black-and-White (0.981 and 0.019 for A and C, respectively) and 138 Jersey (0.812 and 0.188 for A and C, respectively) cows. Preliminary analysis showed no significant associations between PRLR genotypes and milk performance traits. However, Jersey cows of CC genotype produced more milk with higher protein content than those of AA and AC genotypes. Because of the low number of cows of CC genotype, it is necessary to investigate more numerous population of cattle in which all genotypes will be efficiently represented.
This study analyzed the association between single nucleotide polymorphism (A/C) in position -28 located in the TATA box of LTF gene and the lactoferrin concentration in bovine milk secreted by healthy and infected udders. Out of 241, 69 cows were selected into the experimental group and were divided into 3 groups according to mean value of somatic cell count (SCC): I <180 000 cells/mL, II: 180 000 - 350 000 cells/mL and III > 350 000 cells/mL. In each SCC group, three LTF genotypes: AA, AC and CC were identified by PCR-SSCP method. A total of 604 milk samples were collected monthly and lactoferrin concentration was measured by ELISA. The 1-way ANOVA within SCC groups was performed to estimate association of -28 A/C genotypes with mean lactoferrin concentration per lactation. In the group of healthy cows (<180 000 cells/mL) LTF concentration in milk cows with the AA genotype (107.58 ± 17.92 μg/mL) was significantly higher than in homozygotes CC (52.09 ± 19.01 μg/mL). Unexpectedly, in cows with elevated SCC (>350 000 cells/mL) we observed a significant opposite relationship (207.21 ± 28.50 in CC vs 115.0 ± 28.6 μg/mL in AA). We hypothesized that a promoter with allele C, which cannot be recognized as a TATA sequence is becoming more accessible for other transcription factors, which may induce alternative LTF gene expression. We assume that our results demonstrate a very interesting effect of differential gene expression depending on polymorphism in a key regulatory motif (TATA box) and also on the health status of mammary tissues.
Modern pig production needs new tools for fast, reliable, more effective breeding. In the present paper we present a chip containing 45 SNP (Single Nucleotide Polymorphisms) which enables the determining of 1 genetic disease (PSS – Porcine Stress Syndrome), 4 QTLs genes: PRKAG3, CAST, MC4R and ESR, which together with the remaining SNPs create a panel useful in marker-assisted selection and veterinary control. The SNPs were genotyped using the PCR-APEX (Arrayed Primer Extension) technique. Special attention is paid to evaluation of the 45 SNP chip as an alternative approach to parentage and identity control. Based on allele frequency estimations, for a sample of 88 individuals of commercial pig lines, the probabilities that a randomly chosen candidate parent would be excluded from paternity or maternity were estimated to be 99.9% when genotypes of both parents and a progeny were known, and 98% when the genotypes of only one parent and a piglet were available. The marker set presented here also reached a probability of identity in the order of 10-16, which allows for unequivocal discrimination of animals or their products among billions of individuals. Further improvements for upcoming chip versions were also considered.
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