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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. -
Reference genes for livestock gene expression profiling – Literature review
81-89Views:195Quantitative real-time polymerase chain reaction (qPCR) is an essential tool for understanding animal cell’s response to developmental progression or to different experimental conditions at gene expression level. However the reliability of this method heavily lies on proper normalization (measuring a target and a reference gene’s expression from the same sample to correct for technical related variations).
Our literature review aimed to summarize the articles addressing the most important livestock species in regards of reference gene stability used as normalizers for quantitative real-time polymerase chain reaction experiments. Stably expressing reference genes were categorized into 14 distinct groups according to gene function. The number of reference genes tested and the publication numbers according to years and the ranking algorithms were also noted.
Counting showed that genes encoding ribosomal protein components are ranked as most stable in majority of cases and therefore should be taking into account for qPCR stable normalizer gene finding experiments. -
Effects of bioactive plant extracts on immune-related gene expression of common carp (Cyprinus carpio)
49-56Views:308In recent years, intensive fish farming has led to an outbreak of several diseases, and the health status of fish can affect the economy of aquaculture. Since fish health and intestinal health are in correlation, it may also have an impact on immunity. Accordingly, many natural feed additives are being used to improve immune functions. In our study, carotenoids, oligosaccharides, and anthocyanins were applied at 1 m/m% in feed to investigate their effects on cytokines, such as interleukin-1β (IL-1β), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and interferon regulatory factor-1 (IRF-1) in spleen and mid-intestine of 6 months old carp. Gene expression analysis was carried out to examine IL-1β, IL-8, TNF-α, and IRF-1 mRNA levels in fish spleen and mid-intestine. The gene expression level of pro-inflammatory IL-1β decreased in the mid-intestine of carotenoid-fed carp compared to anthocyanin supplemented group, but the effects of the bioactive plant extracts were not observed on the examined cytokines compared to control fish.
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Investigation of intersexual polled goats
11-15Views:179The sex reversal may occur in all mammalian species, but is connected to a favourable trait – the polledness – only in the goat. Later abnormal sex ratio was noticed in these goat populations, in which a part of the phenotypically male individuals was sterile. These males have two X chromosomes. In goats the PIS (Polled Intersex Syndrome) mutation is responsible for the absence of horns in homozygous and heterozygous individuals. This same mutation causes a female-to-male sex reversal, but only in the homozygous polled genetic female goats. The PIS mutation inhibits the expression of the FOXL2 gene which is responsible for ovary development, and a protein encoded by this gene inhibits the activity of the Sox9 gene. The Sox9 gene stimulates the development of the cells of the testis. When the FOXL2 gene is inhibited, the Sox9 gene is activated and transforms the ovary cells into testis cells. In our article we briefly introduce the morphological and chromosome investigations of three intersex individuals we found.
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DHAC-induced transgene overexpression in 35S-gshI GMO poplar (Populus × canescens)
78-83Views:46Relative gene expression levels of transgene gshI (γ-glutamylcysteine synthetase cloned from E. coli) were analyzed by qRT-PCR in two transgenic poplar (Populus × canescens) clones (11ggs and 6lgl) and wild type (WT). An extremely high expression level of transgene gshI was observed in the 6lgl clone (13.5-fold) compared to 11ggs (1.0) samples, which level was doubled (1.8-fold) in the DHAC (5.6-dihydro-5'-azacytidine hydrochloride) treated (at 10-4 M for 7 days) 6lgl samples but not in the 11ggs clone (0.4-fold). Contrary to this result, relative copy number of transgene gshI in the 6lgl clone was found to be less 60% less (1.0) then in the 11ggs samples (1.6). Relative expression levels of proper poplar gene gsh1-poplar showed significantly higher responsiveness to DHAC treatment than transgene gshI with the highest expression level in the untransformed (WT) poplar
clone (19.7-fold) compared to transformed 6lgl (8.7-fold) and 11ggs (2.5-fold) clones. For internal controls constitutively expressed housekeeping genes a-tubulin were applied. For data analysis, the 2−ΔΔCt method was used. DHAC applied in long-term cultures (27 days) at low concentrations (10-8 – 10-6 M) showed morphogenetic activity by initiating de novo root development of leaf discs. -
Isolation of promoters of tissue and ripening-specific strawberry genes by TAIL-PCR and bioinformatic characterization of the sequences
91-99Views:68Isolation of ripening- and tissue-specific promoters has become a very important subject of the genetic regulation and plant physiology research in recent years. It could be possible to reveal the regulation of gene expression, and it may be a very useful approach in the biotechnology. In our work, we have isolated promoter regions of genes exhibiting ripening- and tissuespecific expression in our previous experiments, and the data were
characterized by bioinformatic methods. In the sequence of the ripening-specific Spatula and ACC-oxidase promoters (ACCoxidase is one of the key-enzymes of ethylene biosynthesis, directly related to the process of ripening); we could identify auxin- and ethylene-related cis-regulatory elements. This suggests that there is an interaction between ripening and ethylene-synthesis, in case of non-climacteric strawberry, too. We investigated the promoter regions of three green receptacle-specific genes (putative nitrilaselike protein, Ring transcription factor and an aquaporin protein)
and we could identify several regulatory elements, which refer to hormonal regulation. Additionally, we could find several cisacting elements which associated with stress-responsiveness and endospermium-specific expression. -
Overview of the evolutionary history and the role in citric acid production of alternative oxidase
83-88Views:148All organisms are exposed to countless environmental effects, which influence in a disadvantageous way their life processes. They continuously adapt to the changing conditions and respond to the stress impacts by defence mechanisms. Through different signal transduction pathways they are able to increase or decrease the expression of their genes and consequently modify their metabolic processes. My interest focuses on alternative oxidase (AOX) enzyme whose expression is often increased under biotic and abiotic stress. The so far proven and putative functions of the AOX play a role in the ability of organisms to adapt to different conditions, such as heavy metals accumulation, pathogenic infection, oxidative stress and lack of oxygen or nutrients.
AOX is a member of the di-iron carboxylate protein family. Members of the di-iron carboxylate protein family are present in all kingdoms of life. They are considered to have ancient origin. It is believed that their sulfide-resistant and oxygen-reducing ability played a role in the survival of organisms during the transition between the anaerobic and the aerobic world. It is assumed that the AOX arose in eukaryotes through a primary endosymbiotic event, and this event made possible the development of mitochondria. Afterwords, vertical inheritance, and secondary and tertiary endosimbiotic events led to its spread among eukaryotes. It is assumed that bacteria obtained AOX by horizontal gene transfer from plants.
AOX-catalyzed alternative respiration plays an important role in the operation of energy-producing and biosynthesizing system of microorganisms. In these cases, the regeneration of reduced cofactors is an essential condition, and therefore may be rate-limiting for biotechnological processes, including the citric acid production.
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Studies of Expression of Peptaibol Synthetase of Trichoderma reesei
188-190Views:144Because of the potential importance of peptaibols in the biological control of plant diseases, a transgenic, a T. reesei strain carrying a tex1-promoter: goxA fusion plasmid was constructed for furthur studies. The peptaibol synthetase gene (which is highly similar to T. virens tex1) was identified in the genome sequence of T. reesei. A 900 bp 5’ upstream noncoding fragment, presumed to include the promoter region of tex1, was cloned into the pSJ3 plasmid (which contains the Aspergillus niger goxA gene encoding glucose oxidase). Finally, we transformed T. reesei with the tex1-promoter: goxA fusion containing pSJ3 plasmid.
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Increase of Wheat (Triticum aestivum L.) Resistance to Leaf Rust (Puccinia tritici) via Gene Transformation
127-129Views:81Leaf rust is one of the most significant fungal disease of wheat not only in Hungary but also in other parts of the world. For improving leaf rust resistance of winter wheat variety (Hajdúság, 2003) produced by conventional breeding methods, verified by results of variety tests, showing outstanding results in the aspect of the most important economic values, integration of tissue culture technics, genetic engineering and traditional
methods may provide facilities. Building the gene(s) responsible for resistance into the determined genome can improve the resistance in a way that changes other features of the plant slightly or not at all. In the course of genetical transformation of the variety Hajdúság we applied one of the wheat’s own effecient green-tissue specific insurer genetical regulator, the promoter of ribulose carboxylase 1-5 bisphosphate (RuBisCo) ‘s small
subunit to control the expression of the gene cmg1.