Sung-Jong Oh

Growth hormone-releasing hormone (GHRH) polymorphisms associated with carcass traits of meat in Korean cattle

BackgroundCold carcass weight (CW) and longissimus muscle area (EMA) are the major quantitative traits in beef cattle. In this study, we found several polymorphisms of growth hormone-releasing hormone (GHRH) gene and examined the association of polymorphisms with carcass traits (CW and EMA) in Korean native cattle (Hanwoo).ResultsBy direct DNA sequencing in 24 unrelated Korean cattle, we identified 12 single nucleotide polymorphisms within the 9 kb full gene region, including the 1.5 kb promoter region. Among them, six polymorphic sites were selected for genotyping in our beef cattle (n = 428) and five marker haplotypes (frequency > 0.1) were identified. Statistical analysis revealed that -4241A>T showed significant associations with CW and EMA.ConclusionOur findings suggest that polymorphisms in GHRH might be one of the important genetic factors that influence carcass yield in beef cattle. Sequence variation/haplotype information identified in this study would provide valuable information for the production of a commercial line of beef cattle.

Genetic polymorphisms of the bovine fatty acid binding protein 4 gene are significantly associated with marbling and carcass weight in Hanwoo (Korean Cattle).

The objective of this study was to investigate an association between polymorphisms in the FABP4 gene and phenotypic variation for marbling and carcass weight (CWT) in a population of Hanwoo steers. We re-sequenced 4.3 kb of the FABP4 gene region in 24 Hanwoo bulls and identified 16 SNPs and 1 microsatellite polymorphism. Of these 16 SNPs, three SNPs [g.2774G>C (intron I), g.3473A>T (intron II) and g.3631G>A (exon III, creating a p.Met >Val amino acid substitution)] were genotyped in 583 steers to assess their association with carcass traits. The g.3473A allele showed a significant increasing effect on CWT (P = 0.01) and the g.3631G allele was associated with higher marbling score (P = 0.006). One haplotype of these three SNPs (CAG) was significantly associated with CWT (P = 0.02) and marbling score (P = 0.05) and could potentially be of value for marker assisted selection in Hanwoo cattle. The CAG haplotype effect for CWT was larger (11.14 +/- 5.03 kg) than the largest single locus effect of g.3473A>T (5.01 +/- 2.2 kg).

Use of a bovine genome array to identify new biological pathways for beef marbling in Hanwoo (Korean Cattle)

BackgroundMarbling (intramuscular fat) is a valuable trait that impacts on meat quality and an important factor determining price of beef in the Korean beef market. Animals that are destined for this high marbling market are fed a high concentrate ration for approximately 30 months in the Korean finishing farms. However, this feeding strategy leads to inefficiencies and excessive fat production. This study aimed to identify candidate genes and pathways associated with intramuscular fat deposition on highly divergent marbling phenotypes in adult Hanwoo cattle.ResultsBovine genome array analysis was conducted to detect differentially expressed genes (DEGs) in m. longissimus with divergent marbling phenotype (marbling score 2 to 7). Three data-processing methods (MAS5.0, GCRMA and RMA) were used to test for differential expression (DE). Statistical analysis identified 21 significant transcripts from at least two data-processing methods (P < 0.01). All 21 differentially expressed genes were validated by real-time PCR. Results showed a high concordance in the gene expression fold change between the microarrays and the real time PCR data. Gene Ontology (GO) and pathway analysis demonstrated that some genes (ADAMTS4, CYP51A and SQLE) over expressed in high marbled animals are involved in a protein catabolic process and a cholesterol biosynthesis process. In addition, pathway analysis also revealed that ADAMTS4 is activated by three regulators (IL-17A, TNFα and TGFβ1). QRT-PCR was used to investigate gene expression of these regulators in muscle with divergent intramuscular fat contents. The results demonstrate that ADAMTS4 and TGFβ1 are associated with increasing marbling fat. An ADAMTS4/TGFβ1 pathway seems to be associated with the phenotypic differences between high and low marbled groups.ConclusionsMarbling differences are possibly a function of complex signaling pathway interactions between muscle and fat. These results suggest that ADAMTS4, which is involved in connective tissue degradation, could play a role in an important biological pathway for building up marbling in cattle. Moreover, ADAMTS4 and TGFβ 1could potentially be used as an early biological marker for marbling fat content in the early stages of growth.

Generation of Leukemia Inhibitory Factor-Dependent Induced Pluripotent Stem Cells from the Massachusetts General Hospital Miniature Pig

The generation and application of porcine induced pluripotent stem cells (iPSCs) may enable the testing for safety and efficacy of therapy in the field of human regenerative medicine. Here, the generation of iPSCs from the Massachusetts General Hospital miniature pig (MGH minipig) established for organ transplantation studies is reported. Fibroblasts were isolated from the skin of the ear of a 10-day-old MGH minipig and transduced with a cocktail of six human factors: POU5F1, NANOG, SOX2, C-MYC, KLF4, and LIN28. Two distinct types of iPSCs were generated that were positive for alkaline phosphatase activity, as well as the classical pluripotency markers: Oct4, Nanog, Sox2, and the surface marker Ssea-1. Only one of two porcine iPSC lines differentiated into three germ layers both in vitro and in vivo. Western blot analysis showed that the porcine iPSCs were dependent on LIF or BMP-4 to sustain self-renewal and pluripotency. In conclusion, the results showed that human pluripotent factors could reprogram porcine ear fibroblasts into the pluripotent state. These cells may provide a useful source of cells that could be used for the treatment of degenerative and genetic diseases and agricultural research and application.

The Pig Genome Database (PiGenome): an integrated database for pig genome research

We established the Pig Genome Database (PiGenome) for pig genome research. The PiGenome integrates and analyzes all publicly available genome-wide data on pigs, including UniGenes, sequence tagged sites (STS) markers, quantitative trait loci (QTLs) data, and bacterial artificial chromosome (BAC) contigs. In addition, we produced 69,545 expressed sequence tags (ESTs) from the full-length enriched cDNA libraries of six tissues and 182 BAC contig sequences, which are also included in the database. QTLs, genetic markers, and BAC end-sequencing information were collected from public databases. The full-length enriched EST data were clustered and assembled into unique sequences, contigs, and singletons. The PiGenome provides functional annotation, identification of transcripts, mapping of coding sequences, and SNP information. It also provides an advanced search interface, a disease browser, alternative-splicing events, and a comparative gene map of the pig. A graphical map view and genome browser can map ESTs, contigs, BAC contigs (from the National Institute of Animal Science), Sino-Danish Pig Genome Project transcripts, and UniGene onto pig genome sequences which include our 182 BAC contigs and publically available BAC sequences of the Wellcome Trust Sanger Institute. The PiGenome is accessible at http://pigenome.nabc.go.kr/.

Application of DNA markers in animal industries

The current animal industry is both technology-intensive and globalised. Efficient molecular tools, such as DNA markers, are in demand to strengthen competitive power by maximising the improvement of livestock and obtaining the trust of customers by the verification of product origins. This review describes the present techniques applying DNA markers in the animal industry, with a focus on beef cattle and pigs. Preliminary data from an individual traceability assay for Hanwoo (Korean cattle) using 20 microsatellite markers is described. The potential uses of the assay are demonstrated for several key markers of different traits: for the porcine stress syndrome gene using the RYR mutation; for acid meat using the PRKAG3 mutation; for intramuscular fat using the FABP3 mutation and for fixing the Dominant white allele using KIT duplication. In addition, a possible strategy is suggested to discriminate between pig breeds using mutations of KIT, MC1R, ND2 and the 11-bp insertion in the D-loop of mitochondrial DNA. The industrial application of DNA techniques is limited at present, however, it is expected that DNA markers originating from trait genes, especially those of low-heritability and difficult-to-measure traits, may contribute to maximising the improvement of the major economic traits of animals in the future.

Association Between the Polymorphism on Intron 5 of the Lipoprotein Lipase Gene and Carcass Traits in Hanwoo (Korean cattle)

The primary role of lipoprotein lipase(LPL) is the hydrolysis of triglycerides(TG) from the core of trigly- ceride-rich lipoproteins such as chylomicrons and very low density lipoproteins in plasma. Fatty acids liberated by LPL on capillary endothelial surfaces are available for tissues as energy sources especially in muscles or for storage in the form of TG in adipose tissues. Therefore, as the candidate gene related to the carcass traits of the beef cat- tle, we have directly sequenced the exon 5~exon 6 region in the bovine LPL gene for discovery of single nucleo- tide polymorphism(SNP) with 24 unrelated Hanwoo(Korean cattle). Novel eight sequence variants were detected: three loci on exon 5, three on intron 5 and two on exon 6. All SNPs identified were strongly linked each other, and one hundred twenty eight Hanwoo samples were genotyped one SNP on intron 5 using PCR-restriction fragment length polymorphism method by digestion with Hae III restriction enzyme. The allele frequency of the poly- morphism was 0.76 and 0.24. The effects of this polymorphism on the breeding values of the carcass weight, loin muscle area, back fat thickness and marbling score were analyzed using least square methods of SAS GLM. The marbling score of BB genotype was significantly higher than those of AA and AB genotypes(P < 0.05). This result indicates that this polymorphism may be associated with the variation of marbling score. Further study is warranted to investigate the phenotypic association in Hanwoo.

Detection of Novel Mutations in the FABP3 Promoter Region and Association Analysis with Intramuscular Fat Content in Pigs

Intramuscular fat content(lMF) is considered as one of major economic traits in the pig breeding and industry. In general, high IMF results in better meat quality. Several approaches to detect quantitative trait 10ci( QTL) for IMF indicated a strong possibility of the existence of a QTL related to IMF between the microsatellite marker SW71 and SW1881 on SSC6q. Porcine FABP3 has been considered as a candidate gene affecting IMF due to its physiological roles and position on the pig genome. Two novel mutations, g.-114T> C and g.-158T>G were detected by duplicate sequencing of the porcine FABP3 promoter region. These two mutations were identified as absolute linkage disequilibrium. The g.-158T> G mutation was used for investigating relationships with growth and fat deposition traits. The GG genotype of the g.-158T> G polymorphism showed highly negative effects(P

A Study on Estimation of Individual Growth Curve Parameters and their Relationships with Meat Quality Traits of Crossbred between Korean Native Boars and Landrace Sows

This study was conducted to estimate the growth curve parameters of crossbreds between Korean native boars and Landrace sows and their relationships with meat qualities. The data used were weight-age data and carcass data from 131 males and 122 females raised at N.L.R.I in Korea. Growth curve parameters were estimated from nonlinear regression using Gompertz model individually. Average mature weight(A), average maturing rate(k), and average inflection point( tI ) showing maximum growth rate estimated were 179.54 ± 6.06kg, 0.3154 ± 0.0059, and 5.50 ± 0.11 months in females, and 179.84 ± 6.33kg, 0.3049 ± 0.0061, and 5.24 ± 0.13 months in males, respectively. For the growth curve parameters and derived statistics, the phenotypic correlations of maturing rate with gain rate at inflection, mature weight, and inflection point were .30, .77, and .93 in male, and .31, .78 and .94 in female, respectively. Matrure weight was positively correlated to the inflection point as + .89 in both male and female, indicating that late maturing pigs with lower k had longer maturing period with increasing gain rate and reached point of inflection later than early maturing pigs with higher k, and grew to larger mature weight. Backfat thickness and crude fat contents were correlated with mature weight positively in male and negatively in female, and correlated with gain rate at inflection point positively in both male and female, of which coefficients were as high as .42 and .50 in male, respectively.

Association of Succinate Dehydrogenase and Triose Phosphate Isomerase Gene Expression with Intramuscular Fat Content in Loin Muscle of Korean (Hanwoo) Cattle

In a previous study, succinate dehydrogenase (SDH) and triose phosphate isomerase (TPI) were detected by 2D gel electrophoresis as differentially expressed proteins in the longissimus thoracis muscles of cattle aged between 12 and 27 months old. In the present study, we investigated the association of SDH and TPI gene expression with intramuscular fat content in 50 Hanwoo steers. The SDH gene was expressed at a 4 times higher level in the 12 month old group than in the 27 month old group (p