• Journal of Animal Environmental Science
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• v.3 no.2
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• pp.133-143
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• 1997

Optimal conditions for collecting, storing and drying temperature to utilize slaughter porcine blood for blood meals and the effects of blood meal on growth in broiler chicks were investigated. Dry matter and protein contents of slaughter procine blood were 19.5% and 77%(dry basis), respectively. As for the composites of amino acids in the blood, aspartic acid, arginine, glycine, histidine, leucine, lysine, phenylalanin threonine were shown high. There was no significant difference between the collections by bloodletting and vacuumming in terms of microbial contamination. Storage of slaughter porcine blood showed no differences in protein, DNA and triglyceride contents and pH between the storage methods of freezing (-20$^{\circ}C$) and refrigerating (-4$^{\circ}C$). In case of room temperature storage, however, the decrease in pH and the appearance of new protein due to microbial contaminations increased as the storage periods were prolonged. When drying was done by flash methods, the drying period got shortened as the temperature became higher, yet protein and triglyceride were destoryed more. When drying was done over 120$^{\circ}C$, even at the same degree, the breakdowns of protein and triglyceride increased more as drying period got longer. In feeding trials of broiler chicks, dietary supplementation of the flash dried blood meal at 2% level showed significant difference in growth rate(P＜.05%). These results indicated that the appropriate handling and manufacturing of slaughter porcine blood enabled the blood to be used as a protein source for broiler chicks.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.515-530
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• 1995

Coronaviridae에 속하는 transmissible gastroenteritis virus(TGEV)와 porcine epidemic diarrhea virus(PEDV)를 specific하게 detection할 수 있는 방법을 개발하고자 본 연구를 수행하였다. 두 바이러스 모두 RNA 바이러스이기 때문에 reverse transcription-polymerase chain reaction(RT-PCR)으로 nucleocapsid(N) protein gene의 cDNA를 증폭시켰다. SmaI으로 처리한 pTZ19R에 ligation시킨 후 염기서열을 밝히고자 sequencing하였다. 각각의 prototype virus와 비교하여 상동성을 밝혔다. 두 바이러스에 대한 cDNA probe를 제작하여 Southern blot hybridization을 실시하였다. TGEV의 경우 백신주인 P45와 병독주인 Miller strain을 사용하였다. cDNA를 증폭시키기 위해 N1/N1R과 N2/N2R 두 가지 primer를 이용한 결과, N1/N1R primer의 경우 586bp 크기의 PCR product를 얻을 수 있었고, N2/N2R primers로 582bp의 cDNA를 증폭시킬 수 있었다. PEDV 실험을 위하여 PED 임상 증상을 나타내는 분변을 이용하여 RT-PCR을 실시하였다. P2/P2R primer로 753bp의 PCR product를 얻을 수 있었다. TGEV의 두 가지 strain의 N protein gene을 sequencing하여 prototype인 Purdue strain과 염기서열 상동성을 조사한 결과, 97%이상의 높은 homology를 나타내었다. PED-V 역시 N protein gene을 sequencing하여 CV777과 염기서열 상동성을 조사한 결과 97%이상의 homology로 PEDV임을 알 수 있었다. TGEV와 PEDV의 염기서열을 비교한 결과 29%의 낮은 homology를 관찰할 수 있었다. 두 가지 바이러스의 N protein gene에 대한 cDNA probe를 제작하여 Southern blot hybridization을 한 결과, 각 바이러스에 매우 특이적 반응을 나타내었다.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.191-191
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• 2004

Transgenic animals are produced primarily by microinjecting exogenous DNA into the male pronuclei of a zygote. Microinjection method for gene transmitting is successful in mice but not efficient in farm animals, limiting it's general utility such as a large scale facility and labour. Based on our finding that sperm cells bind with exogenous DNA, sperm was used as a vector for producing transgenic animals to introduced green fluorescence protein(GFP) gene. (omitted)

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.11
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• pp.1572-1575
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• 2008

The xenotransplantation of pig organs and cells can be related with a risk of transmission of infectious diseases to human. Previous findings indicate that the regulatory region of PERV for retroviral transcription, replication and integration into the cellular DNA is located on the 5' Long Terminal Repeat (LTR). The objective of this study is the investigation of methylation and deletion status of the PERV 5' LTR region which can be used for regulating PERV expression. We compared the sequences of genomic DNA and bisulfite-treated genomic DNA from PK-15 cells expressing PERV to observe the methylation status of the 5' LTR. Our results showed that the CpG sites of U3 were methylated and methylation was inconsistent in the R and U5 regions. Also, variable numbers of 18 bp repeats and 21 bp repeats were detected on 5' LTR by sequencing analysis. The consistent U3 methylation might be indicative of host suppression of expression of the retroviruses.

• Korean Journal of Veterinary Service
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• v.30 no.3
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• pp.353-362
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• 2007

A total of 1,354 samples was collected from bovine and porcine carcass from January 2005 to December 2006 in a slaughter house. Twenty five strains(1.8%) of Listeria monocytogenes were isolated from 1,354 samples using selective media. Ten(1.4%) L monocytogenes were isolated from the 677 of bovine carcasses, and 15(2.2%) were isolated from the 677 of porcine carcasses. Among 15 L mono-cytogenes from porcine, 11 siolates were serovars 1/2c, followed by 1/2b (3 strains, 20.0%) and 1/2a(1 strain) Out of 10 bovine samples, positive cases in 1/2a were 9 strains (90.0%), 1/2b were 1 strains(10.0%). PCR primers were selected to amplify a 520-base pair(bp) DNA fragment from the listeolysin O gene (hlyA) of L mono-cytogenes. A 520-bp product was detected in PCR with DNA from L monocytogenes, but not from the other Listeria species tested. A total of 25 L monocytogenes strains were analysed by PFGE after digestion with Apa I. PFGE analysis of genomic DNA showed the $14{\sim}18$ fragments ranging in size from 30 to 550 kb, resulting in 14 patterns.

• Korean Journal of Veterinary Research
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• v.42 no.1
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• pp.65-71
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• 2002

We have utilized the PCR-RFLP method to detect the ryanodine receptor(RYR1) gene mutation and to estimate the genotype frequencies of the RYR1 gene in commercial crossbred pig population. The exon region(659bp) including point mutation(C ${\rightarrow}$T; Arg ${\rightarrow}$Cys) in the porcine ryanodine receptor gene, which is a causal mutation for PSS, was amplified by PCR and digested with Cfo I restriction enzyme. The RYR1 gene was classified into three genotypes by agarose gel electrophoresis. The normal homozygous(NN) individuals showed two DNA fragments consisted of 493 and 166bp. The mutant homozygous(nn) individuals showed only one DNA fragment of 659bp. Also, all three fragments(659, 493 and 166bp) were showed in heterozygous(Nn) carrier animals. The proportions of normal, carrier and PSS pigs within crossbred population of pigs were 81%, 15% and 4%, respectively. According to the results of analysis of variance for the association of genotypes of RYR1 of pigs at 30kg, day age at 90kg and average daily gains, the RYR1 nn genotype was very higher than RYR1 NN genotype for day age at 30kg with 5% level of significant difference, but no significant difference for association of any other genotypes with day age at 90kg and average daily gain in crossbred pigs. Therefore, DNA diagnosis by using PCR-RFLP analysis for the PSS gene was useful for large-scale screening of commercial pigs in the swine industry.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.9
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• pp.1237-1241
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• 2005

Mannose-P-dolichol utilization defect 1 (MPDU1) gene is required for utilization of the mannose donor MPD in synthesis of both lipid-linked oligosaccharides (LLOs) and glycosylphosphatidylinositols (GPI) which are important for functions such as protein folding and membrane anchoring. The full length cDNA of the porcine MPDU1 was determined by in silico cloning and rapid amplification of cDNA ends (RACE). The deduced amino acid showed 91% identity to the corresponding human sequence with five predicted transmembrane regions. RT-PCR was performed to detect its expression pattern in five tissues and results showed that it is expressed ubiquitously among the tissues checked. A single nucleotide substitution resulting in the amino acid change (137 Tyr-137 His) was detected within exon 5. Allele frequencies in six pig breeds showed distinctive differences between those Chinese indigenous pigs breeds and European pigs. Using the pig/rodent somatic cell hybrid panel (SCHP), we mapped the porcine MPDU1 gene to SSC12, which is consistent with the comparative mapping result as conservative syntenic groups presented between human chromosome 17 and pig chromosome 12.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.5
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• pp.768-774
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• 2007

To study the effect of dietary docosahexaenoic acid (DHA) enrichment on the expression of hepatic genes in pigs, weaned, crossbred pigs (30 d old) were fed diets supplemented with either 2% tallow or DHA oil for 18 d. Hepatic mRNA was extracted. Suppression subtractive hybridization was used to explore the hepatic genes that were specifically regulated by dietary DHA enrichment. After subtraction, we observed 288 cDNA fragments differentially expressed in livers from pigs fed either 2% DHA oil or 2% tallow for 18 d. After differential screening, 7 genes were found to be differentially expressed. Serum amyloid A protein 2 (SAA2) was further investigated because of its role in lipid metabolism. Northern analysis indicated that hepatic SAA2 was upregulated by dietary DHA enrichment (p<0.05). In a second experiment, feeding 10% DHA oil for 2d significantly increased the expression of SAA2 (compared to the 10% tallow group; p<0.05). The porcine SAA2 full length cDNA sequence was cloned and the sequence was compared to the human and mouse sequences. The homology of the SAA2 amino acid sequence between pig and human was 73% and between pig and mouse was 62%. There was a considerable difference in SAA2 sequences among these species. Of particular note was a deletion of 8 amino acids, in the pig compared to the human. This fragment is a specific characteristic for the SAA subtype that involved in acute inflammation reaction. Similar to human and mouse, porcine SAA2 was highly expressed in the liver of pigs. It was not detectable in the skeletal muscle, heart muscle, spleen, kidney, lung, and adipose tissue. These data suggest that SAA2 may be involved in mediation of the function of dietary DHA in the liver of the pig, however, the mechanism is not yet clear.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.7
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• pp.1045-1050
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• 2002

The aims of this study were to detect spontaneously occurring apoptosis in cultured porcine ovarian cells, to examine the role of growth hormone (GH), tyrosine kinase (TK), protein kinase G (PKG) and cyclin-dependent kinase (CDK) in the control of this process, and to determine whether the effect of GH on apoptosis is mediated by TK-, PKG- and cdc2-dependent intracellular mechanisms. We studied the action of pGH (10 ng/ml), blockers of TK (genistein, lavendustin, both 100 ng/ml), PKG (Rp-Br-PET-cGMPS, 50 nM; KT5823, 100 ng/ml) and CDK (olomoucine, $1{\mu}g/ml$), as well as combinations of GH with these blockers, on the onset of apoptosis in cultured granulosa cells isolated from antral (3-6 mm) porcine follicles. The functional characteristics of an early apoptotic event, DNA fragmentation, were determined using terminal deoxynucleotidyltransferase (TdT)-mediated dUTP nick end labelling (TUNEL), whilst morphological signs of advanced apoptosis such as pyknosis, chromatin marginalization, shrinkage and fragmentation of nucleus, were detected using routine light microscopy. After culture, some ovarian granulosa cells exhibited DNA fragmentation, which in some cases was associated with morphological apoptosis-related changes (pyknosis, shrinkage and fragmentation of the nucleus). GH significantly reduced the proportion of TUNEL-positive cells. Neither TK nor CDK blockers when given alone, significantly affected the percentage of TUNEL-positive cells although both PKG blockers significantly increased this index. Furthermore, TK and PKG blockers given together with GH, prevented or reversed the inhibitory effect of GH on apoptosis, whilst the CDK blocker olomoucine promoted it. These observations demonstrate apoptosis in porcine ovaries and suggest the involvement of GH, TK, PKG and CDK in the control of this process. They also suggest that the effect of GH on ovarian apoptosis is mediated or regulated by multiple signalling pathways including TK-, PKG- and CDK-dependent intracellular mechanisms.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.7
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• pp.931-937
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• 2006

The full-length cDNA of the porcine SMPX gene was obtained by the rapid amplification of cDNA ends (RACE). The nucleotide sequences and the predicted protein sequences share high sequence identity with both human and mouse. The promoter of SMPX was sequenced and then analyzed to find the promoter binding sites. The reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that SMPX has a high level of expression in heart and skeletal muscle, a very low expression in lung and spleen and no expression in liver, kidney, fat and brain. Moreover, SMPX has a differential expression level in skeletal muscle, the expression in 65-day embryos being higher than other stages. The porcine SMPX was mapped to SSCXp24 by using a somatic cell hybrid panel (SCHP) and was found closely linked to SW1903 using the radiation hybrid panel IMpRH. An A/G single nucleotide polymorphism (PCR-RFLP) in the 3'-untranslated region (3'-UTR) was detected in eight breeds. The analysis of allele frequency distribution showed that introduced pig breeds (Duroc and Large White) have a higher frequency of allele A while in the Chinese indigenous pig breeds (Qingping pig, Lantang pig, YushanBlack pig, Large Black-White pig, Small Meishan) have a higher frequencies of allele G. The association analysis using an experimental population (188 pigs), which included two cross-bred groups and three pure-blood groups, suggested that the SNP genotype was associated with intramuscular fat content.