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EN
The objective of this study was to evaluate an association between the polymorphism of the porcine pituitary-specific transcription factor gene (POU1F1, previously called PIT1) and carcass quality in F2 animals (grandparents: Zlotnicka Spotted boars and Polish Large White sows) being a part of experimental material prepared for a QTL mapping project. The analysis covered a total of 188 F2 offspring of 13 males and 67 females (F1 generation). The RsaI PCR/RFLP polymorphism of the POU1F1 gene was identified and the least squares method was used to evaluate the significance of its effect on the value of carcass quality traits. Three POU1F1/RsaI genotypes were identified in F2 porkers: EE (n=32), EF (n=68) and FF (n=88). Twenty-four carcass quality traits were measured after 24 h of cooling. The POU1F1/RsaI genotype proved to have a significant effect on the following traits: weight of ham bone and bacon including ribs, fat thickness at the lower back (point K3), over the loin, and average fat thickness (mean of five measurements). These results confirm that the POU1F1 gene may be linked to the gene/genes affecting fat deposition in the pig carcass. Moreover, pigs with the EE genotype had a greater loin eye area and showed a higher meat weight and content of carcass than animals of both EF and FF genotypes (unsignificant association), which suggests that a further study is necessary to confirm or exclude the effect of the POU1F1 gene on these traits.
EN
This study was conducted to detect polymorphisms in intron 1 of porcine POU1F1 (POU domain, class 1, transcription factor 1, Pit1, renamed as POU1F1) by comparative sequencing. Within the intron, 23 sites of variation were identified, including 16 single-nucleotide substitutions, 4 single-nucleotide indels, 2 short (3-bp and 17-bp), and one long (313-bp) indels. Several important regulatory motifs were found within the 313-bp indel by in silico analysis. The 313-bp indel was next genotyped in 11 Chinese native pig breeds and 4 western meat-type pig breeds. The appearance of genotypes varied between breeds: among Chinese native breeds, no AA and AB genotypes were found in Tibetan, Lingao, Min, Rongchang, and Songliao Black pigs, no AA genotype was found in Fenjing and Leping Spotted pigs, whereas in Pietrain and Landrace there were no BB genotypes, and all 19 Duroc pigs were AA homozygotes. The western meat-type pigs had high A allele frequencies and the Chinese pigs had more B alleles, except Jianquhai pigs. A positive association of the AA genotype with birth weight was observed in a commercial pig line. This paper demonstrated that the genetic variation in intron 1 of the pig POU1F1 gene was high and these polymorphisms may provide useful makers for QTL analysis.
EN
The aim of this study was to analyse homology in the telomeric region of chromosome 11 in humans and a corresponding fragment in the porcine genome using the FISH technique. The human band specific probe (HSA 11p15) was used for hybridization with pig chromosomes. The obtained results showed strong signals on human chromosome 11p15, as well as pig chromosome 2p17. Some aspects of the similarity between the human and pig chromosome segments have been discussed.
EN
There are two principal applications of transgenic animals. Best known and most advanced application is to use transgenic animals (called bioreactors or molecular farms) for the production of various proteins or biopolymers of medical significance. The second application concerns efforts to improve the productivity traits of breeding animals. We have worked out methods which allow somatic cloning of mammals and gene knockout methods. These methods have been developing very rapidly in recent years and their efficiency will soon be improved to the extent that they will become profitable. For the time being, DNA microinjection with all its disadvantages, remains the principal method of producing transgenic bioreactors. In this paper, the effectiveness of production of transgenic rabbits, goats and pigs with the use of WAP-Fuc gene construct is presented.
EN
The complete coding sequences of 3 porcine genes ? ASPA, NAGA, and HEXA ? were amplified by the reverse transcriptase polymerase chain reaction (RT-PCR) based on the conserved sequence information of the mouse or other mammals and referenced pig ESTs. These 3 novel porcine genes were then deposited in the NCBI database and assigned GeneIDs: 100142661, 100142664 and 100142667. The phylogenetic tree analysis revealed that the porcine ASPA, NAGA, and HEXA all have closer genetic relationships with the ASPA, NAGA, and HEXA of cattle. Tissue expression profile analysis was also carried out and results revealed that swine ASPA, NAGA, and HEXA genes were differentially expressed in various organs, including skeletal muscle, the heart, liver, fat, kidney, lung, and small and large intestines. Our experiment is the first one to establish the foundation for further research on these 3 swine genes.
EN
Although the content of intramuscular fat (IMF) influences significantly meat quality, it can be estimated only after the slaughter of animals. Variants of the H-FABP gene were suggested as candidate genes influencing the variability of IMF. The effect of H-FABP ? HinfI polymorphism on the content of IMF, backfat thickness, the weight and percentages of major meat parts and of the leg in carcass weight was studied in a group of 97 pigs (46 gilts and 51 barrows) of Large White and Landrace breeds using the test of fattening capacity and carcass value. In the set of experimental animals, the frequencies of genes were H = 0.75 ? 0.03 and h = 0.25 ? 0.03. Biometric analyses did not corroborate differences among different H-FABP ? HinfI genotypes and all the traits under study. Only in genotypes HH and Hh the differences between least-square means of phenotypic IMF values under study were close to the limit of significance (P = 0.06).
EN
The matrix metalloproteinase-2 gene (MMP2) was found to be associated with hip structure in pigs. Recently three quantitative trait loci (QTLs) for loin muscle area were found on chromosome 6, to which MMP2 was mapped. In the present study, association analyses were conducted in two pig populations for a single-nucleotide polymorphism (SNP) c.-1959C>G present at a putative promoter region of the MMP2 gene. The association results showed that the animals with the C allele have a significantly larger loin muscle area than that of the animals with the G allele (P < 0.05). To confirm the present results, further functional studies and an additional causative SNP discovery in pigs with various genetic backgrounds are necessary.
EN
The insulin-like growth factor 2 gene (IGF2) has been described in several studies as a candidate gene for meat efficiency in pigs. IGF2 is a member of the growth factors family and has an effect on development of muscle tissue. The effect of IGF2 gene polymorphism on meat efficiency was analysed in a population of 121 Large White pigs. A PCR-based test and RFLP methods were used for detection of genotypes. Allele A, lacking the restriction site, was characterised by the presence of a 0.9-kb fragment. In allele B, the amplimer was cut into a 0.8-kb fragment and some barely detectable fragments. The statistical analysis was carried out according to the General Linear Model procedure. The genotype frequencies observed were: 1.65%, 33.88%, 64.46% for AA, AB and BB genotypes, respectively. There was a significant difference (P 0.05) between the AB and BB genotypes in live weight before the test. A significant association between AB and BB genotypes and body weight before the test was found. No significant difference in other traits of growth and meat efficiency was observed (P > 0.05).
EN
Knowledge of in vivo relationship between the coactivator PPARGC1A and its target genes is very limited, especially in the pig. In this study, a real-time PCR experiment was performed on longissimus dorsi muscle (MLD) and backfat with 10 presumed PPARGC1A downstream target genes, involved in energy and fat metabolism, to identify possible relationships with PPARGC1A mRNA expression in vivo in the pig (n = 20). Except for UCP3 and LPL, a very significant difference in expression was found between MLD and backfat for all genes (P < 0.01). Hierarchical cluster analysis and the significant pairing of mRNA expression data between sampling locations suggested a genetic regulation of the expression of several target genes. A positive correlation with PPARGC1A was found for CPT1B, GLUT4, PDK4, and TFAM (P < 0.0001). A negative correlation was found for UCP2, FABP4, LEP (P < 0.0001), and TNF (P = 0.0071). No significant correlation was detected for UCP3 and LPL. This study provides evidence for a clear difference in mRNA expression of crucial genes in fat and energy metabolism between 2 important tissues. Our data suggest a clear impact of PPARGC1A on energy and lipid metabolism in vivo in the pig, through several of these downstream target genes.
Biotechnologia
|
2006
|
issue 1
103-109
EN
The paper presents issues that shall be considered and elaborated before xenotransplantation reaches clinical stage. In a scope of a research project, such considerations as rules and procedures of a donor selection, including nature of its genetic modifications, must be undertaken. Another problem are regulatory requirements concerning pre-clinical tests on primates. Detailed conditions and requirements that must be fulfilled before clinical trials start are also discussed in the paper. Patient selection criteria as well as relationship with him/her before and after transplantation shall be clarified. A matter of high importance are organisational guidelines and relevant legal aspects of donor animals, breeding and xenotransplantation product harvesting, as well as ethical dimension of these actions.
EN
The study of candidate genes, based on physiological effects, is an important tool to identify genes to be used in marker-assisted selection programs. In this study, a group of halothane gene-free, non-castrated, male Landrace pigs was used to study the association between polymorphisms in the PIT1 (n = 218), GH (n = 213) and GHRH (n = 206) genes and fat thickness, average daily gain, and the EPD (expected progeny difference) for fat thickness, average daily gain, and litter size. These genes are potential candidate markers because of their important physiological effects. The pigs were genotyped by PCR-RFLP, and the statistical model used to analyze the association between genotypes and the traits measured included genotypes as a fixed effect and age and weight as covariates. PIT1 polymorphisms were associated with fat thickness (P = 0.0019), EPD for average daily gain (P = 0.0001) and EPD for fat thickness (P = 0.0001), whereas GH polymorphisms were associated with fat thickness (P = 0.0326) and average daily gain (P = 0.0127), and GHRH polymorphisms were associated with the average daily gain (P = 0.0001) and EPD for fat thickness (P = 0.0004). These results confirmed the potential usefulness of these genes in marker-assisted selection programs for pig breeding.
EN
The complete coding sequences of porcine genes NCF2, BCKDHB and BCKDHA were amplified by using reverse transcriptase polymerase chain reaction (RT-PCR), basing on the conserved coding sequence information of humans or other mammals. These 3 novel porcine genes were then assigned GeneIDs: 100142665, 100142669 and 100142666. The phylogenetic tree analysis revealed that the porcine NCF2, BCKDHB and BCKDHA all are most closely related to the bovine NCF2, BCKDHB and BCKDHA. Tissue expression profile analysis revealed that porcine NCF2, BCKDHB and BCKDHA genes were differentially expressed in tissues, including skeletal muscle, the heart, liver, fat, kidney, lung, small and large intestine.
EN
In the present study we show FISH localization of 4 porcine BAC clones harbouring potential candidate genes for fatness traits: DGAT1 (SSC4p15), PPARA (SSC5p15), ADIPOR1 (SSC10p13) and CREB (SSC15q24). Until now the CREB and ADIPOR1 genes are considered to be monomorphic, DGAT1 is highly polymorphic, while for the PPARA gene only 1 SNP was identified. Assignment of the studied genes in relation to QTL chromosome regions for meat quality in pig chromosomes SSC4, SSC5, SSC10 and SSC15 is discussed.
EN
The present study describes an analysis of genotype and allele distribution at the porcine GH locus among day-10 pig embryos. Embryos were collected post mortem from 6 crossbred (Danish Landrace ? Yorkshire) sows inseminated with mixed Duroc semen and individually frozen for later analysis. After extraction, DNA was subjected to PCR amplification and restriction analysis with Msp I and Hae II enzymes. The genotype frequencies were: Msp I CD 0.17, DD 0.83; and Hae II AA 0.33, AB 0.58; and BB 0.09. The Msp I CC genotype was not found among analysed embryos. To our knowledge, this is the first report on the genotype and allele distribution at the GH locus among early pig embryos.
EN
Myogenic factor 3 (myf - 3) and myogenic factor 5 (myf - 5) are the products of genes: MYOD1 (MYF3) and MYF5, respectively, which belong to the MyoD family. These transcription factors control the processes of myogenesis. The fragments of both the genes comprising exons and promoters were amplified and sequenced. In the 5'UTR region of gene MYOD1, the G302A transition was identified and it is not recognized by any restriction endonuclease. In the promoter region of gene MYF5 we identified three mutations at positions: A65C (PCR-RFLP/AciI); C580T (PCR-RFLP/FokI) and C613T (PCR-RFLP/HinPI). Mutations C580T and C613T were characteristic for Pietrain ? (Polish Large White ? Polish Landrace) crossbred pigs named Torhyb. The C2931T transition, which is not recognized by any restriction enzyme, was identified in exon 3 of gene MYF5. This mutation results in a change of the amino acid sequence (Leu?Pro). The frequency of particular genotypes at the MYOD1 and MYF5 loci proved to be dependent on pig breed. However, Duroc pigs were monomorphic at all the SNPs presented in this study. These SNPs might be analyzed in a further study as probably influencing carcass meatiness.
EN
For 22 carcass traits, we identified 16 QTLs (based on data for pig resource population no. 214, including 180 F2 hybrids of 3 Yorkshire boars and 8 Meishan sows) and mapped them with the use of 39 microsatellite marker loci on chromosomes 4, 6, 7, 8 and 13. Five QTLs were highly significant (P 0.01 at chromosome level): for skin weight (on chromosome 7 at SW1856 and on chromosome 13 at SW1495), skin percentage (on chromosome 7 between SW2155 and SW1856 and on chromosome 13 between SW1495 and SW520), and ratio of leg and butt to carcass (on chromosome 4 at SW1996). The remaining 11 QTLs were significant (P 0.05 at chromosome level): for backfat thickness at shoulder, loin eye width, loin eye height, fat meat weight, lean meat weight, skin weight, bone weight, skin percentage, fat meat percentage, and ratio of lean meat to fat meat. The proportion of phenotypic variance explained by these QTLs ranged from 0.06% (QTL for loin eye width on chromosome 8 between SW1037 and SW1953) to 18.04% (QTL for ratio of lean meat to fat meat on chromosome 7 between SW252 and SW581). Seven of the QTLs reported here are novel.
EN
PIT1 was chosen as a candidate gene to investigate its associations with growth, meat quality and carcass composition traits in the pig. PIT1 is known as the pituitary-specific activator of the growth hormone in several mammals. Furthermore, PIT1 is a positive regulatory factor of prolactin and thyroid-stimulating hormone b. PIT1 is a member of the POU-domain family of genes and is located on porcine chromosome 13. Two informative three-generation families of the University of Hohenheim were used for the presented investigations. The families were based on crosses of the European Wild boar (W) ? Pietrain (P) and Meishan (M) ? Pietrain (P). Each family included 310 F2 animals. A RsaI (PCR) RFLP described by YU et al. (1994) was used for genotyping the animals. Altogether over 50 parameters of growth, meat quality, carcass composition and stress susceptibility were evaluated concerning their associations with RsaI PCR-RFLP. The statistical model of association analyses was used including fixed effects of sex, family, PIT1 genotypes and covariate age at slaughter. Taking the significance level of p < 0.05 as the basis, fourteen traits of growth and carcass composition were associated with PIT1 genotypes in family W ? P. Results from this study suggest that there are contributions of PIT1 gene to variations in the analysed performance traits in pigs. The influence of PIT1 genotypes could not be confirmed under the supposition of a genome-wide test limit.
EN
The ryanodine receptor gene (RYR1) and the estrogen receptor gene (ESR) are the best commercially used markers for predisposition of stress susceptibility (malignant hyperthermia ? MH) and increased litter size, respectively. A simplified method of simultaneous detection of MH and ESR genotypes has been developed. The method is based on simultaneous amplification of fragments of two genes by multiplex PCR and subsequent digestion of the products with two restriction enzymes. The PCR and the digestion could be performed in a single tube and all genotypes could be detected by electrophoretic separation on the same agarose gel. Thus, the development of the method can decrease the cost of the sample analysis and increase the speed and efficiency of the analysis. In our study, frequencies of mutated T allele of the RYR1 gene in Large White (LW), White Meaty (WM) and Landrace (L) were 0.11, 0.13, and 0.15, respectively. Frequencies of the preferred B allele of the ESR gene in the same breeds were 0.35, 0.26, and 0.06, respectively.
EN
It is currently debated whether identification of ESR (estrogen receptor) genotypes should be introduced into breeding programs of Large White pigs. The aim of this study was to evaluate the possible relations between ESR/Ava I polymorphism and carcass performance traits in Polish Large White boars. We examined 103 boars originating from one herd in NE Poland. ESR/Ava I genotypes were determined by the PCR-RFLP method. By the use of the Duncan test, we found highly significant differences (P < 0.01) between WW and MW genotypes, as well as significant differences (P < 0.05) between WW and MM genotypes for meatiness. No significant differences were found for daily gain and selection index.
EN
Few reports exist on complex functions of pig's central nervous system. A direct access to thalamic structures enables a deeper understanding of neuronal networks. Here we present an easy to implement stereotactic approach to reach both reticular and dorsolateral thalamic nuclei (RTN and LD). In thirteen pigs (7 weeks old) the correct electrode position was confirmed for 22 out of 26 thalamic electrodes (RTN: A+2, L9, V24 and LD: A-2, L5, V20, with bregma A 0, L 0). Quantitative effects of isoflurane/nitrous oxide (State 1) and fentanyl sedation (State 2) were determined by brain hemodynamics and metabolism. Neurophysiologic features were performed by spectral power, coherence and SEP analysis. Brain blood flow (by 21 ? 13%) and oxidative brain metabolism (CMRO2 by 26 ? 12%, CMRGlucose by 26 ? 22%) were markedly reduced during State 1 (P<0.05). Regional thalamic blood flow exhibited similar alterations, but side-differences did not occur. State 1 induced quite similar brain activity in cortical as well as thalamic regions investigated. During State 2 electrocortical activity of low frequency ranges was markedly reduced, whereas spectral band power of high frequency ranges was additionally decreased in RTN (P<0.05). Thus, we used a convenient approach for targeted deep electrode implementation and characterized electrophysiological features in RTN and LD.
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