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1

New SNPs in the coding and 5' flanking regions of porcine MYOD1 (MYF3) and MYF5 genes

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Urbanski P. , Kuryl J.

Journal of Applied Genetics

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2004

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tom 45

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nr 3

325-329

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.

2

Radiation Hybrid Panel--modern method for physical mapping of porcine genome

100%

Korwin-Kossakowska A. , Kuryl J.

Biotechnologia

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2004

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nr 3

184-192

EN

The creation of high-resolution maps is central to genetics: maps allow genes to be associated with phenotypes and can be used as tools for gene cloning. Maps for the human genome and other mammalian species are being created both by classical recombination methods and by physical mapping techniques. One powerful method for creating physical maps is the use of irradiation and fusion gene transfer (IFGT) to make radiation hybrid maps. Goss and Harris who showed that chromosome fragments, generated by lethal irradiation of donor human cells, could be 'rescued' by fusion to rodent recipient cells originally developed this technology. Each resulting hybrid cell, which includes both hamster and human DNA, can be grown up to yield a hybrid cells or hybrid cell line of cells containing the same random subset of the human genome. A radiation hybrid panel (RH) consists of a number of different hybrid cell lines (sometimes just called hybrids or clones). The recombinant clone is tested for the presence or absence of each DNA marker (for example microsatellites). By estimating the frequency of breakage, and thus the distance between markers, it is possible to determine their order along chromosome. The successful use of RH panels to map the human genome has led to the development of WG-RH (whole genome radiation hybrid) maps in other species, for example of the porcine genome. This map provides a resource for rapid, large scale physical mapping of the swine genome and facilitates resolving genetic and physical distances prior to designing strategies for positional candidate cloning of the gene(s) contributing to economically important traits. WG-RH panels have now been used to create chromosomal maps in several species: rat, horse, bovine, cat, dog and mouse.

3

A novel polymorphism in exon 1 of the porcine myogenin gene

100%

Wyszynska-Koko J. , Kuryl J.

Journal of Applied Genetics

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2005

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tom 46

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nr 4

399-402

EN

Myogenin is a gene belonging to the MyoD family, which codes for the bHLH transcription factor playing a key role in myogenesis. It affects the processes of differentiation and maturation of myotubes during embryogenesis. Fragments of the porcine myogenin coding sequence and promoter region were amplified and subjected to MSSCP analysis. TC transition recognised by the MaeIII restriction enzyme in exon 1 was revealed, which appeared to be a silent mutation in the region of the transactivation domain. No other polymorphism was found either in the remaining coding sequence or the promoter region.

4

Association of POU1F1/RsaI genotypes with carcass traits in pigs

80%

Kuryl J. , Pierzchala P. M. , Pierzchala M.

Journal of Applied Genetics

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2001

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tom 42

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nr 3

309-316

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.

5

Evaluation of the genetic profile of the Pulawska breed

80%

Babicz M. , Kuryl J. , WalkiewiczA.

Journal of Applied Genetics

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2003

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tom 44

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nr 4

497-508

EN

An assessment was made of the genetic variation of the Pulawska pig through the determination of polymorphism of 6 genes and 14 microsatellite sequences. The examinations covered 52 gilts included in a preservation breeding project. The identification of the alleles at microsatellite loci was performed in an ABI PRISM 310 GENETIC ANALYZER. Gene polymorphism was established by the PCR-RLFP method. On the basis of the variation of 6 genes and 14 microsatellites the mean value of the heterozygosity coefficient was estimated at 0.61, while the value of the corresponding PIC coefficient (polymorphism information content) amounted to 0.55. The probability that the genotypes of two randomly chosen individuals in a population are identical was: 6.95 ? 10?3 (based on gene allele frequency) and 1.23 ? 10?14 (based on microsatellite allele frequency).

6

A new SNP in the promoter region of the porcine MYF5 gene has no effect on its transcript level in m. longissimus dorsi

70%

Urbanski P. , Flisikowski K. , Starzynski R. R. , Kuryl J. , Kamyczek M.

Journal of Applied Genetics

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2006

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tom 47

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nr 1

59-61

EN

Myogenic factor 5 (myf-5) is the product of the MYF5 gene, belonging to the MyoD family. This transcription factor participates in the control of myogenesis. We identified 3 new mutations in the promoter region of the gene: A65C, C580T and C613T. The aim of this study was to evaluate the influence of the A65C transversion on gene expression. The analysis was conducted on 15 Polish Large White gilts. The relative content of MYF5 mRNA in m. longissimus dorsi did not differ significantly across MYF5 genotypes (AA, AC, CC). This result suggests that the A65C transversion may not play an important role in the expression of the MYF5 gene in analysed adult muscle but it abolishes a putative binding site for two transcription factors (CDP and HSF1) and creates such a site for Sp1.

7

Polymorphism of the porcine growth hormone gene and its linkage to hormone gene and its linkage to microsatellites S0083 and S0090

61%

Korwin-Kossakowska A. , Pierzchala M. , Kuryl J. , Zwierzchowski L. , Cymerowska-Prokopczyk I. , SiadkowskaE.

Journal of Applied Genetics

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1999

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tom 40

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nr 2

85-91

EN

Polymorphism of microsatellites S0083 and S0090 as well as of the second exon and second intron of the pGH (porcine growth hormone) gene was determined for 293 pedigrees obtained from crossing of 12 F1 (Zlotnicka Spotted x Polish Large White) boars with 64 F1 (Zlotnicka Spotted x Polish Large White) sows, experimental material arranged for a QTL mapping project. Microsatellites were genotyped by capillary electrophoresis of PCR products in an ABI PRISM 310 Genetic Analyser PERKIN-ELMER. The PCR-RFLP polymorphism of the GH gene was identified using HaeII and MspI restriction endonucleases. The following order of linked loci was established: GH-S0083-S0090. This is important information for the mapping of QTLs on chromosome 12.

8

Polymorphisms in coding and regulatory regions of the porcine MYF6 and MYOG genes and expression of the MYF6 gene in m. longissimus dorsi versus productive traits in pigs

61%

Wyszynska-Koko J. , Pierzchala M. , Flisikowski K. , Kamyczek M. , Rozycki M. , Kuryl J.

Journal of Applied Genetics

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2006

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tom 47

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nr 2

131-138

EN

MYOG and MYF6 belong to the MyoD gene family. They code for the bHLH transcription factors playing a key role in later stages of myogenesis: differentiation and maturation of myotubes. Three SNPs in porcine MYF6 and two in porcine MYOG were analysed in order to establish associations with chosen carcass quality and growth rate traits in Polish Landrace, Polish Large White and line 990 sows. No statistically significant effect of SNP in the promoter region of the MYF6 gene on its expression measured on mRNA level was found. Associations between the genotype at the MYF6 locus and carcass quality traits appeared to be breed-dependent. The C allele in the case of SNP in the promoter region and GC haplotype in exon 1 were advantageous for right carcass side weight in Polish Landrace sows and disadvantageous for this trait in Polish Large White sows. These gene variants were also the most advantageous for loin and ham weight in sows of line 990. The mutation in exon 1 of the MYOG gene had no statistically significant association with carcass quality traits and the mutation in the 3'-flanking region had the breed-dependent effect as well. These results suggest that SNPs analysed in this study are not causative mutations, but can be considered as markers of some other, still unrevealed genetic polymorphism that influences the physiological processes and phenotypic traits considered in this study.