• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.37-43
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• 2004

This study was carried out to investigate PSS (Porcine Stress Syndrome) with the PSE (Pale, Soft, Exudative) in 319 different pigs(Yorkshire 150; Landrace 89 and Duroc 80). The PCR-RFLP method was adapted to detect the ryanodine receptor (RYR 1) gene mutation and to estimate the genotype frequency of the RYR1 gene in breeding pig population. The DNA samples were collected from hair follicles of pigs of Yorkshire, Landrace and Duroc. After DNA amplification by PCR, the PCR products were digested by restriction enzyme, Cfo I. Primary PCR products of ryanodine receptor gene were length of 659 bp in hair follicle and their second PCR products were length of 522 bp in hair follicle. The exon region (522 bp) including point mutation ($C \arrow T; Arg \arrow Cys$) in the porcine ryanodine receptor gene, which is a causal mutation for PSS, was digested with Cfo I restriction enzyme. The RYR1 gene was classifed into three genotypes by agarose gel electrophoresis. The normal homozygous (NN) individuals showed two DNA fragments consisted of 439 and 83 bp. The mutant homozygous (nn) individuals showed only one DNA fragment 522 bp. In addition, all three fragments (522, 439 and 83 bp) were showed in heterozygous (Nn) carrier animals. The normal homozygous (NN), heterozygous (Nn) and mutant homozygous (nn) were 98.00, 2.00 and 0.00% in Yorkshire pigs, 87.64, 11.24 and 1.12% in Landrace, 100.00, 0.00 and 0.00% in Duroc, respectively. The gene frequencies of N and n were 0.990 and 0.010 in Yorkshire pigs, 0.933 and 0.067 in Landrace, 1.000 and 0.000 in Duroc, respectively.

• Analytical Science and Technology
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• v.13 no.4
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• pp.504-510
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• 2000

The impurity profiles from the raw materials of glycyrrhizin were performed by the high performance liquid chromatography (HPLC)/electrospray ionization (ESI)- mass spectrometry (MS). For the HPLC experiment, a $C_{18}$($3.9{\times}300mm$, $10{\mu}m$) column was used and the mobile phase was acetic acid/$H_2O$ (1:10):acetonitrile=3:2 with a flow rate of 0.8 ml/min. The effluent was splitted into the ratio of 50:1 and went into the ESI-MS. Three to six impurities were found and informed of the identification of the structure of the impurities by ESI-MS. And the structures of impurities were suggested to a hydroxy-glycyrrhizin which is added with hydroxy group (-OH) in the glycyrrhetic acid moiety and a reduced-glycyrrhizin which the position of 12 of the glycyrrhetic acid moiety is reduced. The purities of the standard materials were about 90%.

• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.155-164
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• 2004

This involves identifying and cloning trapped genes from cultured cells carrying the gene-trap constructs and generating cloned zebrafish using these cells for functional study. Gene-trapping studies in gene-trapped cells were carried out in initial and cloned zebrafish carrying gene-trap events were successfully produced based on the nuclear transplantation technique. Two kind of retroviral gene-trap constructs were adopted. The first one(SA/GFP-TP), constructed in my laboratory, carries a GFP reporter gene containing a splicing acceptor and an internal neo gene. The second one(Neo-TP), obtained from Dr. Hicks (Hicks et al., 1997), contains a promoter-less neo gene located in the LTR sequence of a retroviral vector. The infected cells were subjected to drug selection(neomycin treatment) because the two constructs carry the neomycin resistant gene. All those cells survived the neomycin treatment should carry the proviral insertions. For Neo-TP, Isolated DNA from the neomycin-resistant fibroblast cells infected by Neo-TP, was digested with EcoR1 restriction enzyme and transformed into bacteria after ligation. This procedure led to the isolation of seven clones carrying flanking cellular DNA with a typical retroviral integration signature sequence. These clones contained genomic DNA ranging from 1kb to 7kb and sequences of 300-600 bp were obtained from each of the rescued plasmids. Database searching showed that all of them share high homology to zebrafish sequences. For fish cloning using tagged cells, initially, nucleus donors directly selected from a mixture of cells(Neo-TP cells) were used. A total of 44 embryos(3.7%) out of 1179 transplants were reached blastula stage; 8 of these embryos(0.8%) hatched and 3(0.3%) of them survived to adulthood. One out of three lived cloned zebrafish has an amplified fragment and was labeled with 32P.

• Restorative Dentistry and Endodontics
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• v.20 no.2
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• pp.645-654
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• 1995

Porphyromonas endodontalis is a black-pigmented anaerobic Gram negative rod which is associated with endodontal infections. It has been isolated from infected dental root canals and submucous abscesses of endodontal origin. DNA probe is an available alternative, offering the direct detection of a specific microorganism. Nucleic-acid probes can be off different types: whole different: whole-genomic, cloned or oligonucleotide probes. Wholegenomic probes are the most sensitive because the entire genome is used for possible hybridization sites. However, as genetically similar species of bacteria are likely to be present in specimences, cross-reactions need to be considered. Cloned probes are isolated sequences of DNA that do not show cross-reactivity and are produced in quantity by cloning in a plasmid vector. Cloned probes can approach the sensitivity found with whole-genomic probes while avoiding known cross-reacting species. Porphyromonas endodontalis ATCC 35406 (serotype $O_1K_1$) was selected in this experiment to develop specific cloned DNA probes. EcoR I-digested genomic DNA fragments of P. endodontalis ATCC 35406 were cloned into pUC18 plasmid vector. From the E. coli transformed with the recombinant plasmid 4 clones were selected to be tested as specific DNA probes. Restriction-digested whole-genomic DNAs prepared from P. gingivalis 38(serotype a), W50(serotype b), A7A1-28(serotype C), P. intermedia 9336(serotype b), G8-9K-3(serotype C), P. endodontalis ATCC 35406(serotype $O_1K_1$), A. a Y4(serotype b), 75(serotype a), 67(serotype c), were each seperated on agarose gel electrophoresis, blotted on nylon membranes, and were hybridized with digoxigenin-dUTP labeled probe. The results were as follows: 1. Three clones of 1.6kb(probe e), 1.6kb(probe f), and 0.9kb(probe h) in size, were obtained. These clones were identified to be a part of the genomic DNA of P. endodontalis ATCC 35406 judging from their specific hybridization to the genomic DNA fragments of their own size on Southern blot. 2. The clones of 4.9kb(probe i) was identified to be a part of the genomic DNA of P. endodontalis ATCC 35406. but not to specific for itself. It was hybridized to P. gingivalis A7A1-28, P. intermedia G89K-3.

• Restorative Dentistry and Endodontics
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• v.20 no.2
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• pp.399-422
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• 1995

The purpose of this study was to investigate a fragment of possibility of pulpotomy with the Nd-YAG laser by the observation of pulpal healing process and the fine structural changes of the fibroblasts of the remaining pulpal tissues. Class V cavities on !55 teeth from 4 adult dogs were prepared and the pulp chambers were opened with a sterilized round bur. In the control group(19 teeth), the exposed coronal pulps were excised by a sharp excarvator. After bleeding was controlled with the sterilized cotton pellets, calcium hydroxide powder was applied on the remaining pulpal tissues and the cavities were sealed with Z.O.E. In the experimental group 1 : the pulpotomy with laser-calcium hydroxide powder application group(l9 teeth), the exposed coronal pulps were excised by Nd-YAG laser(10 watts power, 2 psi water, 20 psi air) for 2 or 3 seconds and calcium hydroxide powder was applied on the remaining pulpal tissues and the cavities were sealed with Z.O.E. In the experimental group 2 : the pulpotomy with laser-no calcium hydroxide powder application group(17 teeth), after amputating the coronal pulps with Nd-YAG laser as the experimental group 1, the remaining pulpal tissues were covered with stenilized aluminum foil and the cavities were filled with Z.O.E. The animals were sacrificed at the intervals of 1, 2, 3 and 4 weeks. All the teeth were rouutinely processed and the remaining pulpal tissues were observed by the light microscope and electron microscope. The results were as follows : 1. In light microscopic findings, there was no significant difference of the inflammatory response in the remaining pulpal tissues between the control group and the experimental groups. In both of the experimental group 1 : pulpotomy with laser-calcium hydroxide powder application group and the control group, the dentin bridges were observed after 2 weeks and the structure of the dentin bridge was almost same. In the experimental group 2 : pulpotomy with laser-no calcium hydroxide powder application group, the fibrous layers instead of dentin bridge were observed on the superficial portion of the remaining pulpal tissues after 2 weeks and they were consisted with densely crowded active fibroblasts. 2. In the electronmicroscopic findings, the active fibroblasts in the experimental groups were more frequently observed than in the control group at 1 week. But active fibroblasts were found with same frequency after 2 weeks in all of the control group and the experimental groups. 3. General distortions of the cell such as loss of the cell membrane, vaculoization of the cell etc. were observed at the suberficial layer of the remaining pulpal tissues and the carbonization was found in the dentinal wall in 1 week of the experimental groups. 4. In the experimental group 2 : pulpotomy with laser-no calcium hydroxide powder application group, the activity and the density of the fibroblasts in the fibrous layer were more than those in the deep portion of the remaining pulpal tissues after 2 weeks. 5. In the control group, bacteria such as cocci and bacilli were observed frequently, but in the experimntal groups, they could not be observed.

• Applied Biological Chemistry
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• v.40 no.5
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• pp.380-387
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• 1997

Phytopathogenic Erwinia carotovora subsp. carotovora (Ecc) LY34 causes plant tissue maceration by secretion of pectinolytic enzymes such as pectate Iyase (PL) existed as multiple isoenzyme form. Genomic DNA from Ecc LY34 was digested with Sau3Al and ligated into the BamHI site of pBluescript ll $SK^+$. Among them, a clone hydrolyzing polypectate was selected and its DNA was digested with BamHI. Through the subsequent subcloning the resulting 3.1 kb fragment, corresponding to a peICI, was subcloned into pLYPA 100. The structural organization of a peICI gene encoding a 374 amino acid residues consists of an open reading frame (ORF) of 1,122 bp commencing with a ATG start codon and followed by a TAA stop codon. PeICI contained a typical prokaryotic signal peptide of 22-amino acid. Since the deduced amino acid sequences of PeICl protein was very similar to those of PelIII of Erwinia carotovora subsp. carotovora, and to those of Pel3 of Erwinia carotovora subsp. atroseptica, and to those of PeIC of Erwinia carotovora subsp. carotovora, it belong to the same family PLbc group. The 374-amino acld PeICI had a calculated Mr of 40,507 and pI of 7.60.

• Applied Biological Chemistry
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• v.39 no.3
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• pp.195-199
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• 1996

We have cloned several E. coli sfs genes which stimulate mal gene expression with $crp^{{\ast}1}$). One the genes (pPVC2) was sequenced and potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. In order to investigate the regulation of the sfs1 gene by the cAMP-CRP complex, we have constructed the sfs-lacZ fusion gene in this research. The overall transcriptional stimulations of sfs1 gene in the presence cAMP were confirmed by ${\beta}-galactosidase$ activity and Western blot analysis of sfs1-lacZ fusion gene. Transcriptional regulation by cAMP-CRP was also confirmed by Northern blot analysis. End-labelled DNA of the DNA fragment in sfs1 regulation region were used for gel retardation assay to examine the CRP-DNA complex in the presence of cAMP. Results here indicate that CRP binding site in the regulatory region of sfs1 gene is positive regulator for the expression of sfs1 gene.

• Applied Biological Chemistry
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• v.47 no.2
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• pp.182-186
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• 2004

Morphological observation of roots and molecular technique were used to investigate the symbiotic relationships between arbuscular mycorrhizal (AM) fungi and ginseng roots. Korean ginseng, Panax ginseng, was collected from 8 sites in Korea. Colonization pattern of AM fungi in ginseng roots was determined as an Arum type under light microscopes. Nested PCR using AM fungal specific primers was employed to amplify a partial region on 18s rDNA of AM fungi from the root extracted mixed DNA. The amplified DNA was cloned and analyzed by random fragment length polymorphism (RFLP) with restriction enzymes, AluI, HinfI and AsuC21. One from each RFLP pattern was selected for sequencing. A total 16 clones were sequenced and identified as 2 species of AM fungi; Paraglomus brasilianum and Glomus spurcum. Paramglomus brasilianum was found from most of the ginseng roots, in this syudy suggesting that this species of AM fungi could have specific relationship with the ginseng root. Possible roles of AM fungal species in ginseng roots are discussed.

• Journal of KIISE:Computing Practices and Letters
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• v.10 no.6
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• pp.484-498
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• 2004

In this paper. we propose a novel performance monitoring scheme for heterogeneous SOAP nodes. The scheme is basically based on two-level (kernel-level and user-level) packet filtering of TCP flows. By TCP flow, we mean a sequence of raw packet streams on a TCP transaction. In this scheme, we detect and extract SOAP operations embedded in SOAP messages from TCP flows. Therefore, it becomes possible to monitor heterogeneous SOAP nodes deployed on diverse SOAP-based middlewares such as .Net and Apache AXIS. We present two implementation mechanisms for the proposed scheme. The first mechanism tries to identify SOAP operations by analyzing all fragmented SOAP messages on TCP flows. However, a naive policy would incur untolerable overhead since it needs to copy all packets from kernel to user space. The second mechanism overcomes this problem by selectively copying packets from kernel to user space. For selective copying, we use a kernel-level packet filtering method that makes use of some representative TCP flags.(e.g. SIN, FIN and PSH). In this mechanism, we can detect SOAP operations only from the last fragment of SOAP messages in most cases. Finally, we implement a SOAP monitoring system using a component ca]led SOAP Sniffer that realizes our proposed scheme, and show experimental results. We strongly believe that our system will play a vital role as a tool for various services such as transaction monitoring and load balancing among heterogeneous SOAP nodes.

• Research in Plant Disease
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• v.14 no.1
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• pp.71-75
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• 2008

An isolate of Peanut stunt virus (PSV), named as Tr-PSV, was isolated from white clover (Trifolium repens L) showing mosaic symptom. Tr-PSV systemically infected all plants tested in the Nicotiana spp. and induced local lesions on inoculated leaves of Chenopodium amaranticolor. However, Tr-PSV induced typical mosaic symptoms as ER-PSV on Vigna unguiculata 5 to 6 days after inoculation, while Fny-CMV used as a control virus of Cucumovirus produced local lesions on inoculated leaves. In dsRNA analysis, Tr-PSV consisted of four dsRNAs, but satellite RNA was not detected. The cDNA of coat protein gene of Tr-PSV was amplified by RT-PCR using a Cucumovirus-specific single pair primers that designed to amplify a DNA fragment of approximately 950 bp. By restriction mapping analysis using RFLP of the RT-PCR products and by serological properties of gel diffusion test, Tr-PSV belongs to a typical member of PSV subgroup I. This is the first report on the occurrence of PSV in white clover in Korea.