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We generated the PEDE (Pig EST Data Explorer; http://pede.dna.affrc.go.jp/) database using sequences assembled from porcine 5' ESTs from oligo-capped full-length cDNA libraries. Thus far we have performed EST analysis of various organs (thymus, spleen, uterus, lung, liver, ovary and peripheral blood mononuclear cells) and assembled 68,076 high-quality sequences into 5546 contigs and 28,461 singlets. PEDE provides a search interface for getting results of homology searches and enables users to obtain information on sequence data and cDNA clones of interest. Single-nucleotide polymorphisms detected through comparison of the EST sequences are classified by origin (western and oriental breeds) and are searchable in the database. This database system can accelerate analyses of livestock traits and yields information that can lead to new applications in pigs as model systems for medical research.

The development of informative polymorphic markers is essential for QTL mapping. We developed 50 microsatellite markers from BAC clones containing genes that were predicted to map swine chromosome 4 (SSC4) according to comparative analysis between human and swine chromosomes, and constructed a linkage map that consisted of 37 markers including 24 markers closely linked to genes in BAC clones. Microsatellite markers were developed by direct-sequencing of BAC clones and our results demonstrated that this method was effective for developing microsatellite markers in specific regions on chromosomes. Effective development of microsatellite markers closely linked to genes can further accelerate the comparative studies of chromosomes between different species.

Loci affecting swine intramuscular fat content, backfat thickness, carcass weight, and daily weight gain were assigned to regions of swine chromosome (SSC) 4, which were shown to correspond to human chromosome (HSA) 1p22--> q25 by ZOO-FISH, bidirectional chromosome painting, as well as by the linkage map of genes. In order to select candidate genes responsible for the above traits from the human genome database, precise correspondence between SSC4 and HSA1 is a prerequisite. In the present study, 27 genes, PTGFR, GBP1, GBP2, GFI1, GCLM, ABCD3, EXTL2, KCNA3, ADORA3, KCND3, WNT2B, NRAS, SYCP1, PTGFRN, IGSF2, NOTCH2, S100A10, SHC1, SSR2, LMNA, CCT3, CD5L, PEA15, FCER1G, EAT2, DDR2, and LAMB3, located in the HSA1 region corresponding to SSC4 or possibly SSC4, were assigned to the IMpRH map. The alignment of genes from centromere to telomere in the SSC4 q arm is basically conserved in HSA1p22-->q25 with the direction from the q arm to the p arm, which is in good agreement with results from linkage mapping. In addition, the present study first demonstrated that WNT2B residing in the middle of the HSA1 region was assigned to SSC18 with a high lod score (> 5), and that at least three intrachromosomal rearrangements occurred in the region in the process of swine and human evolution. PTGFR, and LAMB3 localized at both ends of the HSA1 region were assigned to SSC6 and SSC9, respectively, which is consistent with regional correspondence reported earlier. In the course of the above analysis, microsatellite markers were developed in the proximity of eleven genes localized on SSC4, and three genes on other swine chromosomes., (Copyright 2003 S. Karger AG, Basel)

Interleukin 7 (IL7) is a cytokine that has many immunological functions, including regulation of hematopoiesis and peripheral lymphocytes. cDNA and a genomic DNA segment containing the porcine IL7 gene were isolated and sequenced, showing that porcine IL7 consists of 176 amino acids and that its gene spans over about 13 kb of genomic DNA. Porcine IL7 has 85% and 73% homology with human IL7 in terms of the nucleotide and amino acid sequences, respectively. Whereas the murine IL7 gene does not have an exon corresponding to human exon 5 (Lupton et al., 1990), the porcine IL7 gene was found to contain the same exon-intron structure as the human gene. These findings, together with the upstream structure of the cDNA elucidated in the present study, indicate that the relationship between swine and human IL7 is closer than that between mouse and human IL7. The IL7 gene was mapped to swine chromosome 4q11-->q13 by fluorescence in situ hybridization and, using a radiation hybrid panel, was localized between microsatellite markers Sw1336 and Sw1073 on the same chromosome., (Copyright 2001 S. Karger AG, Basel)

The pig aminoacylase 1 (ACY1; N-acylamino acid aminohydrolase) gene was isolated from a pig cosmid library and characterized. The gene spans about 4.7 kb and consists of 15 exons. Fluorescence in situ hybridization found ACY1 to be located on pig chromosome 13 in the region q21-->q22. This result and previous reports show that a large part of pig chromosome 13 corresponds to human chromosome 3. BLAST search results reveal that chromosome 13 contains a transcript similar to human ribosomal protein L29 (RPL29)/heparan sulfate/heparin-interacting protein (HIP). The transcript lies near the 3'-flanking region of the pig ACY1 gene; the 2 genes are linked tail-to-tail. The deduced amino acid sequence shows distinct homology to human RPL29/HIP, 96% identical in the N-terminal region. In the corresponding human 3p21.1 region, a deletion closely linked to the ACY1 locus has been observed in carcinoma cells. This suggests that a tumor suppressor gene is located in this region. Comparative mapping suggests also that the human RPL29/HIP gene may be near ACY1. Because many growth factors and cytokines interact with cells via heparin/heparan sulfate-proteoglycan, RPL29/HIP may play an important role on the cell surface by modulating interactions between cells and extracellular molecules.

We found a unique exon-intron structure of the porcine S100C gene, a member of the S100 family, in which all other genes characterized to date have common exon-intron organization. The genomic DNA encoding the porcine S100C was cloned and the entire nucleotide sequence of the gene was analyzed. The gene is present as a single copy and consists of three exons and two introns with a total size of 5.3 kb. The first intron is located after the ATG translation initiation codon. Such an intron has never been found in other S100 genes, but is found in almost all calmodulin genes. The gene structural similarity suggests a close evolutionary relationship between the S100C gene and calmodulin genes.