Rapid Communication: The correlation between histone modifications and expression of key genes involved in accumulation of adipose tissue in the pig.

Histone modification is a well-known epigenetic mechanism involved in regulation of gene expression; however, it has been poorly studied in adipose tissues of the pig. Understanding the molecular background of adipose tissue development and function is essential for improving production efficiency and meat quality. The objective of this study was to identify the association between histone modification and the transcript level of genes important for lipid droplet formation and metabolism. Histone modifications at the promoter regions of 6 genes (, , , , , and ) were analyzed using a chromatin immunoprecipitation assay. Two modifications involved in activation of gene expression (acetylation of H3 histone at lysine 9 and methylation of H3 histone at lysine 4) as well as methylation of H3 histone at lysine 27, which is known to be related to gene repression, were examined. The level of histone modification was compared with transcript abundance determined using real-time PCR in tissue samples (subcutaneous fat, visceral fat, and longissimus dorsi muscle) derived from 3 pig breeds significantly differing in fatness traits (Polish Large White, Duroc, and Pietrain). Transcript levels were found to be correlated with histone modifications characteristic to active loci in 4 of 6 genes. A positive correlation between histone H3 lysine 9 acetylation modification and the transcript level of ( = 0.53, < 4.8 × 10), ( = 0.34, < 0.02), and ( = 0.43, < 1.0 × 10) genes was observed. The histone H3 lysine 4 trimethylation modification correlated with transcripts of ( = 0.64, < 4.6 × 10) and ( = 0.37, < 0.01) genes. No correlation was found between transcript level of all studied genes and histone H3 lysine 27 trimethylation level. This is the first study on histone modifications in porcine adipose tissues. We confirmed the relationship between histone modifications and expression of key genes for adipose tissue accumulation in the pig. Epigenetic modulation of the transcriptional profile of these genes (e.g., through nutritional factors) may improve porcine fatness traits in future.

KCNJ11 and KCNQ1 Gene Polymorphisms Are Not Associated with Post-Transplant Diabetes Mellitus in Kidney Allograft Recipients Treated with Tacrolimus.

Post-transplant diabetes mellitus (PTDM) is a metabolic disorder occurring after solid organ transplantation during the therapy with calcineurin inhibitors. ATP-sensitive potassium channels KCNJ11 and KCNQ1 play an important role in the regulation of insulin secretion by ß cells and development of diabetes mellitus. Numerous studies have confirmed the association between KCNJ11 and KCNQ1 gene polymorphisms and type 2 diabetes. The aim of this study was to examine the association between KCNJ11 and KCNQ1 gene polymorphisms and posttransplant diabetes mellitus in kidney allograft recipients treated with tacrolimus. The study included 201 patients who received kidney transplants. The patients were subdivided into two subgroups: patients with PTDM (N = 35) and patients without PTDM (N = 166). The association between KCNJ11 and KCNQ1 gene polymorphisms and post-transplant diabetes was studied in three models of univariate Cox regression analysis, i.e., additive, dominant and recessive. In these three models there were no statistically significant associations between KCNJ11 and KCNQ1 gene polymorphisms and PTDM. The results of this study suggest lack of association between KCNJ11 and KCNQ1 gene polymorphisms and post-transplant diabetes mellitus in kidney allograft recipients treated with tacrolimus in the Polish population.

Deep sequencing of a candidate region harboring the SOX9 gene for the canine XX disorder of sex development.

A disorder of sex development (DSD) in dogs with female sex chromosomes (78, XX), a lack of the SRY gene and the presence of testes or ovotestes is commonly diagnosed in numerous breeds. The molecular background of DSD is not fully recognized but has been linked to the copy number variation in the region harboring the SOX9 gene. We applied a genome-wide association study and targeted next-generation sequencing techniques to compare DSD and normal female dogs. The genome-wide association study did not indicate a significant chromosome region. Targeted next-generation sequencing of a 1.5-Mb region on canine chromosome 9 harboring the SOX9 gene revealed two putatively DSD-associated copy number variations 355 kb upstream and 691 kb downstream of SOX9, four blocks of low polymorphism and two blocks of an elevated heterozygosity. An initial next-generation sequencing analysis showed an association with two SNPs, but validation in larger cohorts did not confirm this result. We identified a large homologous fragment (over 243.8 kb), named hfMAGI2, located upstream of SOX9, that overlaps a known copy number variation region. It shows a high sequence similarity with the 5' flanking region of the MAGI2 gene located on canine chromosome 18 that encodes a protein involved in ovary formation during early embryonic development. Our study showed that the identified copy number variation region located upstream of the SOX9 gene contains potential regulatory sequences (long non-coding RNA and hfMAGI2) and led to the assumption that a multiplication of this element may alter expression of the SOX9 gene, triggering the DSD phenotype.

FOXO1 is a TXN- and p300-dependent sensor and effector of oxidative stress in diffuse large B-cell lymphomas characterized by increased oxidative metabolism.

Molecular profiling has led to identification of subtypes of diffuse large B-cell lymphomas (DLBCLs) differing in terms of oncogenic signaling and metabolic programs. The OxPhos-DLBCL subtype is characterized by enhanced mitochondrial oxidative phosphorylation. As increased oxidative metabolism leads to overproduction of potentially toxic reactive oxygen species (ROS), we sought to identify mechanisms responsible for adaptation of OxPhos cells to these conditions. Herein, we describe a mechanism involving the FOXO1-TXN-p300 redox-dependent circuit protecting OxPhos-DLBCL cells from ROS toxicity. We identify a BCL6-dependent transcriptional mechanism leading to relative TXN overexpression in OxPhos cells. We found that OxPhos cells lacking TXN were uniformly more sensitive to ROS and doxorubicin than control cells. Consistent with this, the overall survival of patients with high TXN mRNA expression, treated with doxorubicin-containing regimens, is significantly shorter than of those with low TXN mRNA expression. TXN overexpression curtails p300-mediated FOXO1 acetylation and its nuclear translocation in response to oxidative stress, thus attenuating FOXO1 transcriptional activity toward genes involved in apoptosis and cell cycle inhibition. We also demonstrate that FOXO1 knockdown in cells with silenced TXN expression markedly reduces ROS-induced apoptosis, indicating that FOXO1 is the major sensor and effector of oxidative stress in OxPhos-DLBCLs. These data highlight dynamic, context-dependent modulation of FOXO1 tumor-suppressor functions via acetylation and reveal potentially targetable vulnerabilities in these DLBCLs.

Novel polymorphisms in porcine 3'UTR of the leptin gene, including a rare variant within target sequence for MIR-9 gene in Duroc breed, not associated with production traits.

The leptin gene (LEP) is considered as a functional candidate for production traits in livestock due to its crucial role in energy homeostasis. Because polymorphisms in regulatory sequences may affect gene expression, we searched for them in the 3'UTR of LEP and analyzed their association with production traits. Four breeds and a composite line were studied. In the Polish Landrace and Polish Large White breeds, 8 SNPs and 1 indel were observed; whereas, in the Duroc breed, 9 specific SNPs were found. Pietrain and Line 990 were monomorphic. One SNP (g.+168C>T), observed in the Duroc breed only, was located within a target site for microRNA (miR-9). Association studies showed a weak association between one SNP (c.+846C>T) and abdominal fat weight in the Polish Landrace only. Thus, we concluded that contribution of polymorphisms in the 3'UTR to phenotypic variability of pig production traits is marginal. Moreover, we presented an overview of known polymorphisms (128) in the pig leptin gene.

Polymorphism in 3' untranslated region of the pig PPARA gene influences its transcript level and is associated with adipose tissue accumulation.

The PPARA (peroxisome proliferator-activated receptor-α) gene encodes a nuclear receptor that plays an important role in fatty acid catabolism by transcriptional regulation of genes involved in fatty acid oxidation and can be considered as a candidate gene for fatness traits in the pig. The aim of the study was to search for a functional polymorphism in 3' untranslated region (UTR), their association with production traits, and postnatal PPARA transcript level in 2 skeletal muscles (longissimus and semimembranosus) of 5 commercial pig breeds (Polish Landrace [PL], Polish Large White [PLW], Duroc, Pietrain, and Pulawska). Altogether, 9 novel polymorphisms (8 SNP and 1 indel) were found in the 3' UTR. The in silico analysis revealed 6 putative microRNA target sequences in the analyzed region. The c.*636A>G substitution was widely distributed across breeds and located near the putative target sequence for miR-224. The relative PPARA transcript level was higher (P < 0.05) in LM of AA than in those of GG homozygous animals for SNP c.*636A>G. The luciferase assay revealed that miR-224 probably acts as a negative regulator of the PPARA expression in pig adipocytes (P = 2.9 × 10(-7)), but we did not observe the effect of the A or G alleles on the interaction between miR-224 and its putative target sequence. We hypothesize that the 2 predominant haplotypes, differing at 4 sites (including c.*636A>G), present different architecture of its 3' UTR and it could affect the level of the transcript. The c.*636A>G SNP, analyzed in PL and PLW, was significantly associated with backfat thickness at 3 points (P < 0.05) and intramuscular fat content (P < 0.01) in PL. Suggestive associations were found between 4 SNP (c.*321A>C, c.*324G>C, c.*626T>C, and c.*636A>G) and fatty acid contents in LM and subcutaneous and visceral fat tissue of PL, PLW, Duroc and Pietrain pigs. The PPARA mRNA level was higher in semimembranosus muscle than in LM (P = 8.38 × 10(-12)) in a general comparison and the same trend was found in most breeds (except for PL) and at all tested days of age (60, 90, 120, 150, 180, and 210 d). The effect of breed was highly significant in a general comparison (P = 0.48 × 10(-8)), but there was no common expression pattern in both muscles among different age groups. We conclude that the c.*636A>G SNP in the PPARA gene can be considered in PL breed as a useful genetic marker for adipose tissue accumulation.

Transcript level of the porcine ME1 gene is affected by SNP in its 3'UTR, which is also associated with subcutaneous fat thickness.

Pork quality depends on multiple factors, including fatty acid composition in muscle and fat tissues. The ME1 gene is a strong candidate for fat accumulation, as it encodes the malic enzyme, which is required for fatty acid synthesis. We identified seven new polymorphisms in 3'UTR of the ME1 gene and moreover confirmed the presence of 4 polymorphisms detected previously. Interestingly, the studied Duroc pigs were monomorphic at all these polymorphic sites, while in 3 other breeds (Pietrain, Polish Landrace and Polish Large White), the polymorphisms were unevenly distributed. One of the novel SNPs (c.*488A>G) was found in the Polish Large White and the Polish Landrace only, and the association studies revealed that it was significantly associated with backfat thickness and average daily weight gain in the Polish Landrace (N = 207) and the Polish Large White (N = 157). This SNP was differently associated with ME1 transcript level in muscle and backfat. The in silico analysis of another novel SNP (c.*548C>T) indicated that it is located within a binding sequence conserved among vertebrates for the miR-30 family in 3'UTR of the ME1. It was shown that in the longissimus muscle, but not in adipose tissue, CT gilts compared with CC ones had significantly lower levels of the ME1 transcript. This polymorphism, however, was not associated with production traits. Additionally, we observed that transcript level of the ME1 was significantly higher in subcutaneous fat than in the longissimus muscle, as well as both investigated tissues of the Polish Landrace when compared to the other breeds. However, no association was found between this polymorphism and fatty acid profiles. We conclude that the ME1 gene polymorphism (c.*488A>G) is a potential marker for porcine backfat thickness.

Transcript abundance of the pig stearoyl-CoA desaturase gene has no effect on fatty acid composition in muscle and fat tissues, but its polymorphism within the putative microRNA target site is associated with daily body weight gain and feed conversion ratio.

Fatty acid composition in porcine intramuscular fat affects the dietetic value and technological properties of meat. The stearoyl-CoA desaturase (SCD) gene is a strong positional and functional candidate for fatty acid composition. Our sequence analysis in 4 breeds (Duroc, Pietrain, Polish Landrace, and Polish Large White) revealed a novel SNP in the 5'-flanking sequence and 9 novel SNP and 2 novel indels in the 3' untranslated region (UTR). Transcript level of the SCD in subcutaneous fat was significantly greater than in muscle tissue (n=83; P<0.001) and the interbreed comparison revealed a greater transcript level in the fat tissue of Polish Landrace (P<0.01). We found no association between the abundance of the SCD transcript and fatty acid composition in any of the tissues. We performed an association analysis between 4 SNP (c.-353C>T, c.-233T>C, c.*164A>G, and c.*928G>C), 1 indel (c.*2574_2576delGTC), and production traits in Polish Large White (n=185) and synthetic line 990 (n=243). The most pronounced associations were observed for the c.*928G>C polymorphism, which occurs within a predicted target site for 2 microRNA (ssc-miR-185 and ssc-miR-491). In line 990, this polymorphism was significantly associated with daily BW gain (P<0.04 under the general model) and feed conversion ratio (P<0.0004) but not with fatness traits. The same tendency, but not significant, was observed in the Polish Large White breed. When both breeds were analyzed together, these associations were again highly significant (daily BW gain P<0.003; feed conversion ratio P<0.0001). We conclude that c.*928G>C is a promising marker for both porcine traits.

Polymorphisms in the promoter region of the adiponectin (ADIPOQ) gene are presumably associated with transcription level and carcass traits in pigs.

The main goal of this study was to screen for polymorphisms in the porcine adiponectin (ADIPOQ) gene promoter, analyse their influence on transcription and identify any association with production traits in pigs. A 1018-bp region of the ADIPOQ gene promoter was analysed in 113 pigs, and seven novel polymorphisms found. Luciferase assays were performed in HEK293 (human embryonic kidney) cells and primary porcine adipose mesenchymal stem cells (pADMSCs) to investigate their affect on promoter activity. A 16-bp indel (c.-106_-91delGCCAGGGGTGTGAGCC) was found to influence promoter strength in vitro. In the HEK293 cell line, the Del/Del genotype showed greater luciferase activity than did the Ins/Ins genotype (P < 0.01). In pADMSCs, the insertion genotype of the ADIPOQ promoter showed greater luciferase activity than did the deletion genotype (P < 0.01). An association study performed for two novel polymorphisms, c.-67G>A and the 16-bp indel, showed significant correlation with loin measurements in Polish Landrace (P < 0.05) and synthetic line 990 (P < 0.01) pigs.

Two candidate genes (FTO and INSIG2) for fat accumulation in four canids: chromosome mapping, gene polymorphisms and association studies of body and skin weight of red foxes.

Fat accumulation is a polygenic trait which has a significant impact on human health and animal production. Obesity is also an increasingly serious problem in dog breeding. The FTO and INSIG2 are considered as candidate genes associated with predisposition for human obesity. In this report we present a comparative genomic analysis of these 2 genes in 4 species belonging to the family Canidae - the dog and 3 species which are kept in captivity for fur production, i.e. red fox, arctic fox and Chinese raccoon dog. We cytogenetically mapped these 2 loci by FISH and compared the entire coding sequence of INSIG2 and a fragment of the coding sequence of FTO. The FTO gene was assigned to the following chromosomes: CFA2q25 (dog), VVU2q21 (red fox), ALA8q25 (arctic fox) and NPP10q24-25 (Chinese raccoon dog), while the INSIG2 was mapped to CFA19q17, VVU5p14, ALA24q15 and NPP9q22, respectively. Altogether, 29 SNPs were identified (16 in INSIG2 and 13 in FTO) and among them 2 were missense substitutions in the dog (23C/T, Thr>Met in the FTO gene and 40C/A, Arg>Ser in INSIG2). The distribution of these 2 SNPs was studied in 14 dog breeds. Two synonymous SNPs, one in the FTO gene (-28T>C in the 5'-flanking region) and one in the INSIG2 (10175C>T in intron 2), were used for the association studies in red foxes (n = 390) and suggestive evidence was observed for their association with body weight (FTO, p < 0.08) and weight of raw skin (INSIG2, p < 0.05). These associations indicate that both genes are potential candidates for growth or adipose tissue accumulation in canids. We also suggest that the 2 missense substitutions found in dogs should be studied in terms of genetic predisposition to obesity.

Association of MC3R gene polymorphisms with body weight in the red fox and comparative gene organization in four canids.

There are five genes encoding melanocortin receptors. Among canids, the genes have mainly been studied in the dog (MC1R, MC2R and MC4R). The MC4R gene has also been analysed in the red fox. In this report, we present a study of chromosome localization, comparative sequence analysis and polymorphism of the MC3R gene in the dog, red fox, arctic fox and Chinese raccoon dog. The gene was localized by FISH to the following chromosome: 24q24-25 in the dog, 14p16 in the red fox, 18q13 in the arctic fox and NPP4p15 in the Chinese raccoon dog. A high identity level of the MC3R gene sequences was observed among the species, ranging from 96.0% (red fox--Chinese raccoon dog) to 99.5% (red fox--arctic fox). Altogether, eight polymorphic sites were found in the red fox, six in the Chinese raccoon dog and two in the dog, while the arctic fox appeared to be monomorphic. In addition, association of several polymorphisms with body weight was analysed in red foxes (the number of genotyped animals ranged from 319 to 379). Two polymorphisms in the red fox, i.e. a silent substitution c.957A>C and c.*185C>T in the 3'-flanking sequence, showed a significant association (P < 0.01) with body weight.

Estimating myostatin gene effect on milk performance traits using estimated gene content for a large number of non-genotyped cows.

The objective of this study was to estimate the myostatin (mh) gene's effect on milk, protein and fat yield in a large heterogeneous cow population, of which only a small portion was genotyped. For this purpose, a total of 13 992 889 test-day records derived from 799 778 cows were available. The mh gene effect was estimated via BLUP using a multi-lactation, multi-trait random regression test-day model with an additional fixed regression on mh gene content. As only 1416 animals, (of which 1183 cows had test-day records) were genotyped, more animals of additional breeds with assumed known genotype were added to estimate the genotype (gene content) of the remaining cows more reliably. This was carried out using the conventional pedigree information between genotyped animals and their non-genotyped relatives. Applying this rule, mean estimated gene content over all cows with test-day records was 0.104, showing that most cows were homozygous +/+. In contrast, when gene content estimation was only based on genotyped animals, mean estimated gene content over all cows with test-day records was with 1.349 overestimated. Therefore, the applied method for gene content estimation in large populations needs additional genotype assumptions about additional animals representing genetic diversity when the breed composition in the complete population is heterogeneous and only a few animals from predominantly one breed are genotyped. Concerning allele substitution effects for one copy of the 'mh' gene variant, significant decreases of -76.1 kg milk, -3.6 kg fat and -2.8 kg protein/lactation were obtained on average when gene content estimation was additionally based on animals with assumed known genotype. Based on this result, knowledge of the mh genotypes and their effects has the potential to improve milk performance traits in cattle.

New method to combine molecular and pedigree relationships.

Relationship coefficients are traditionally based on pedigree data. Today, with the development of molecular techniques, they are often completely replaced by coefficients calculated from molecular data. Examples are relationships from microsatellites for biodiversity studies but also genomic relationships from SNP as currently used in genomic prediction of breeding values. There are, however, many situations in which optimal combination of both sources would be the best solutions. Obviously, this is the case for incompletely genotyped populations, but also when pedigree information is sparse. Also, markers, even dense ones, do not reflect the whole genome and therefore give only an incomplete picture of relationships. The main objective of this study was therefore to develop a method to calculate a relationship matrix by the combination of molecular and pedigree data. It will be useful for all situations where pedigree and molecular data are available. In this study, based on simulations of pedigree and marker data, we used partial least squares regression and linear regression to combine total allelic relationship coefficients calculated for each marker with additive relationship coefficients calculated from incomplete pedigree. The results showed that the greatest advantage of this method, compared with the one that replaces a part of the pedigree-based relationship matrix by a genomic relationship matrix, is that adding the partial pedigree data allows for the correction of the molecular coefficient for the ungenotyped part of the genome.

A robust method for simultaneous estimation of single gene and polygenic effects in dairy cows using externally estimated breeding values as prior information.

The aim of this study was to develop a robust method to estimate single gene and random polygenic animal effects simultaneously in a small field dataset with limited pedigree information. The new method was based on a Bayesian approach using additional prior information on the distribution of externally estimated breeding values. The field dataset consisted of 40,269 test-day records for milk performance traits for 1455 genotyped dairy cows for the 11 bp-deletion in the coding sequence of the myostatin gene. For all traits, estimated additive effects of the favoured wild-type allele ('+' allele) were smaller when applying the new method in comparison with the application of a conventional mixed inheritance test-day model. Dominance effects of the myostatin gene showed the same behaviour but were generally lower than additive effects. Robustness of methods was tested using a data-splitting technique, based on the correlation of estimated breeding values from two samples, with one-half of the data eliminated randomly from the first sample and the remaining data eliminated from the second sample. Results for 100 replicates showed that the correlation between split datasets when prior information included was higher than the conventional method. The new method led to more robust estimations for genetic effects and therefore has potential for use when only a small number of genotyped animals with field data and limited pedigree information are available.

Postnatal transcription profile and polymorphism of the ADIPOR1 gene in five pig breeds.

As a result of its role in energy homeostasis regulation, the ADIPOR1 gene is a candidate for fat deposition, an important production trait, in the pig. The aim of the study was to conduct a comparative analysis of the ADIPOR1 postnatal transcript level, in order to establish its promoter and 5'UTR sequences and to search the gene for polymorphisms. The transcription level was examined in longissimus dorsi and semimembranosus muscles collected from 180 pigs at 60-210 days of age, representing five pig breeds: Duroc, Polish Large White, Polish Landrace, Pietrain and Pulawska. We calculated highly significant breed by age by muscle interaction (P < 0.0001) and breed by muscle interactions (P < 0.01). The 5'UTR and promoter region of the porcine ADIPOR1 gene were amplified for the first time and their sequences were deposited in the GenBank database. In total, 21 novel and two previously described polymorphisms were found in the ADIPOR1 promoter, coding, intronic, 5' and 3' untranslated regions. The only SNP detected in the coding region was a synonymous substitution. Two polymorphisms in 3'UTR (c.*129A>C and c.*536A>G) showed no significant effect on the transcript level. Our results showed a high polymorphism of the ADIPOR1 and a complexity in its transcription level in the studied muscles. This complexity indicates that conclusions based on such studies should be carefully gradated.

The pig CART (cocaine- and amphetamine-regulated transcript) gene and association of its microsatellite polymorphism with production traits.

The cocaine- and amphetamine-regulated transcript (CART) gene is a candidate gene that may affect performance and body composition traits in the pig. The purpose of this study was to establish the chromosomal localization and genomic sequence of the porcine CART gene, search for polymorphism and analyse its phenotypic effect in 644 pigs representing two breeds, Polish Large White (PLW) and Polish Landrace (PL), and a synthetic line 990 (L990). The CART gene was fluorescence in situ hybridization (FISH)-mapped to the chromosome 16q21. The 1878 bp DNA fragment covering three exons, two introns and the 5' flanking region was sequenced and analysed. A new A/G single nucleotide polymorphism (SNP) at position -238 bp was found. The coding sequence was conserved between porcine and human CART genes. Previously unknown short tandem repeat polymorphism (CA)(2)(CG)(n)(CA)(n) was identified in intron 2. Three alleles 251, 253 and 259 bp were found. The 251-bp allele was predominant in all the analysed populations of pigs, whereas the 253-bp allele occurred with the lowest frequency. The statistical analysis revealed significant allelic additive effects on meat content in carcass (p < 0.05) and abdominal fat weight (p < 0.01) in PLW, and meat content in carcass (p < 0.05) and backfat thickness (p < 0.05) in PL. Our study confirmed that chromosome region harbouring the CART gene is a promising quantitative trait loci for pig production traits.

Sampling genotype configurations in a large complex pedigree.

Many genetic problems can be solved by Monte Carlo method. This often requires sampling genotype configurations over pedigree. Current available samplers are inefficient for large animal pedigrees. A new sampler suitable for large complex pedigrees has been developed and evaluated. The sampler uses simple and iterative peeling algorithms alternately. The sampler was compared to two other samplers on hypothetical pedigree of 79 individuals and recessive disease. The behaviour of the sampler was evaluated in four experimental designs on real bovine pedigree of 907,903 animals. The application of the sampler was also exemplified in identical by descent study.

Accuracy of prediction of gene content in large animal populations and its use for candidate gene detection and genetic evaluation.

To estimate and to use the effects of single genes on quantitative traits, genotypes need to be known. However, in large animal populations, the majority of animals are not genotyped. These missing genotypes have to be estimated. However, currently used methods are impractical for large pedigrees. An alternative method to estimate missing gene content, defined as the number of copies of a particular allele, was recently developed. In this study, the proposed method was tested by assessing its accuracy in estimation and use of gene content in large animal populations. This was done for the bovine transmembrane growth hormone receptor and its effects on first-lactation milk, fat, and protein test-day yields and somatic cell score in Holstein cows. Estimated gene substitution effects of replacing a copy of the phenylalanine-coding allele with a copy of the tyrosine-coding allele were 295 g/d for milk, -8.14 g/d for fat, -1.83 g/d for protein, and -0.022/d for somatic cell score. However, only the gene substitution effect for milk was found to be significant. The accuracy of the estimated effects was evaluated by simulations and permutations. To validate the use of predicted gene content in a mixed inheritance model, a cross-validation study was done. The model with an additional regression of milk, fat, and protein yields and SCS on predicted gene content showed a better capacity to predict breeding values for milk, fat, and protein. Given these results, the estimation and use of allelic effects using this method proved functional and accurate.

Association of a new SNP in promoter region of the porcine FABP3 gene with fatness traits in a polish synthetic line.

Associations between FABP3 (alternatively named H-FABP) gene polymorphisms and fatness traits were tested in two pig breeds (Polish Large White and Polish Landrace) and one synthetic line - 990. Three known single nucleotide polymorphisms, detected by HinfI, MspI and HaeIII restriction enzymes, were analyzed. Moreover, three new polymorphisms in the 5' regulatory region were identified: C(-221)T, C(-160)G and T(-158)G, but only the third one was widely distributed and correlated with backfat thickness in line 990. The obtained results suggest that the FABP3 gene is linked with an unknown gene directly affecting backfat thickness, but the analyzed polymorphisms do not influence fatness traits.