• Tuberculosis and Respiratory Diseases
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• v.49 no.5
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• pp.546-557
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• 2000

Background : Oligonucleotide chip technology has proven to be a very useful tool in the rapid diagnosis of infectious disease. Rifampin resistance is considered as a useful marker of multidrug-resistance in tuberculosis. Mutations in the rpoB gene coding $\beta$ subunit of RNA polymerase represent the main mechanism of rifampin resistance. The purpose of this study was to develop a diagnosis kit using oligonucleotide chip for the rapid and accurate detection of rifampin-resistance in Mycobacterium tuberculosis. Method : The sequence specific probes for mutations in the rpoB gene were designed and spotted onto the glass slide, oligonucleotide chip. 38 clinical isolates of Mycobacterium were tested. A part of rpoB was amplified, labelled, and hybridized on the oligonucleotide chip with probes. Results were analyzed with a laser scanner. Direct sequencing was done to verify the results. Result : The low-density oligonucleotide chip design어 to determine the specific mutations in the rpoB gene of M. tuberculosis accurately detected rifampin resistance associated with mutations in 28 clinical isolates. Mutations at codons 531, 526, and 513 were confirmed by direct sequencing analysis. Conclusion : Mutant detection using oligonucleotide chip technology is a reliable and useful diagnostic tool for the detection of multidrug-resistance in M. tuberculosis.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.11
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• pp.1644-1651
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• 2014

Transgenic animals have become important tools for the production of therapeutic proteins in the domestic animal. Production efficiencies of transgenic animals by conventional methods as microinjection and retrovirus vector methods are low, and the foreign gene expression levels are also low because of their random integration in the host genome. In this study, we investigated the homologous recombination on the porcine ${\beta}$-casein gene locus using a knock-in vector for the ${\beta}$-casein gene locus. We developed the knock-in vector on the porcine ${\beta}$-casein gene locus and isolated knock-in fibroblast for nuclear transfer. The knock-in vector consisted of the neomycin resistance gene (neo) as a positive selectable marker gene, diphtheria toxin-A gene as negative selection marker, and 5' arm and 3' arm from the porcine ${\beta}$-casein gene. The secretion of enhanced green fluorescent protein (EGFP) was more easily detected in the cell culture media than it was by western blot analysis of cell extract of the HC11 mouse mammary epithelial cells transfected with EGFP knock-in vector. These results indicated that a knock-in system using ${\beta}$-casein gene induced high expression of transgene by the gene regulatory sequence of endogenous ${\beta}$-casein gene. These fibroblasts may be used to produce transgenic pigs for the production of therapeutic proteins via the mammary glands.

• The korean journal of orthodontics
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• v.32 no.6 s.95
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• pp.425-434
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• 2002

The purpose of this investigation was to study the spatial changes of the maxillofacial complex following maxillary protraction transmitted to the center of resistance of a dry juvenile human skull by a modified maxillary protraction appliance. Four dry juvenile human skulls (without mandible) with well aligned upper deciduous dentition and early mixed dentition were used as experimental samples. A modified protraction headgear was fabricated from a Delare's facemask, and following an alginate impression, an orthodontic resin maxillary splint was made for each dry skull. Protraction force level was maintained at approximately 1000gm per side for 6 hours. Cephalometric radiographs were taken pre- and post- protraction, and nine reference markers with 1.5 mm length of $.017\times.025$ TMA wire were placed on the right side of the skull for an accurate superimposition of serial cephalometric radiographs. The present investigation demonstrated that vertical changes associated with an anterior displacement of the maxillary complex was observed, and the most prominent effect of protraction headgear was a counterclockwise rotation of the maxilla, that is, a forward and downward tipping around the palatomaxillary region.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.6
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• pp.419-423
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• 2003

Bacterial blight caused by Xantomonas oryzae pv. oryzae is one of the most serious diseases of rice especially in southern area of Korea. Three races, $\textrm{K}_1$, $\textrm{K}_2$ and $\textrm{K}_3$, are the most dominant species. lo improve rice breeding efficiency using marker assisted selection, some RFLP markers were surveyed for polymorphism between resistant and susceptible to $\textrm{K}_1$ and $\textrm{K}_3$. And, 127 doubled-haploid (DH) lines derived from Milyang121/HRl1650-1-4-2 and 131 DH lines derived from Milyang123/HR10624-AC5 were evaluated to bacterial blight ($\textrm{K}_1$ and $\textrm{K}_3$). Milyang121 and HR10624-AC5 have Xa-1, resistant to $\textrm{K}_1$ race, and Milyang123 has Xa-3, resistant to $\textrm{K}_1$ and $\textrm{K}_3$ race. Three markers, RZ590, RZ536 and RG303, showing polymorphism between parents and resistance gene, Xa-1 and Xa-3, were analysed in the two combinations of DH lines. The segregation pattern of resistant DH population of Milyang123/HR10624-AC5 to susceptible showed 3:1 and 1:1 in $\textrm{K}_1$ and $\textrm{K}_3$ race. In three RFLP markers, RZ590 was linked to Xa-1 on chromosome 4, and RZ536 and RG303 were linked to Xa-3 on chromosome 11. The map distance between Xa-1 and RZ590 was 3.1cM on chromosome 4, and Xa-3 and RZ536/RG303 were 7.6/16.0cM on chromosome 11, respectively. The results of RFLP mapping will be useful for the selection and pyramiding of bacterial blight resistant genes.

• Journal of Life Science
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• v.16 no.3 s.76
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• pp.454-458
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• 2006

HSF1 is the heat shock transcription factor in Saccharomyces cerevisiae. S. cerevisiae KNU5377 can ferment at high temperature such as $40^{\b{o}}C$. We have been the subjects of intense study because Hsf1p mediates gene expression not only to heat shock, but to a variety of cellular and environmental stress challenges. Basing these facts, we firstly tried to construct the hsf1 gene-deleted mutant. PCR-method for fast production of gene disruption cassette was introduced in a thermotolerant yeast S. cerevisiae KNU5377, which allowed the addition of short flanking homology region as short as 45 bp suffice to mediate homologous recombination to kanMX module. Such a cassette is composed of linking genomic DNA of target gene to the selectable marker kanMX4 that confers geneticin (G418) resistance in yeast. That module is extensively used for PCR-based gene replacement of target gene in the laboratory strains. We describe here the generation of hsf1 gene disruption construction using PCR product of selectable marker with primers that provide homology to the hsf1 gene following separation of haploid strain in wild type yeast S. cerevisiae KNU5377. Yeast deletion overview containing replace cassette module, deletion mutant construction and strain confirmation in this study used Saccharomyces Genome Deletion Project (http:://www-sequence.standard.edu/group/yeast_deletion_project). This mutant by genetic manipulation of wild type yeast KNU5377 strain will provide a good system for analyzing the research of the molecular biology underlying their physiology and metabolic process under fermentation and improvement of their fermentative properties.

• Horticultural Science & Technology
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• v.19 no.1
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• pp.17-21
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• 2001

To develop a selection system for regenerating plants from transformed tissues, effects of four antibiotics (kanamycin, hygromycin, carbenicillin, cefotaxime) and herbicide (phosphinotricin) on shoot regeneration from cotyledon and hypocotyl explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis) were studied. For cotyledon, shoot induction was not significantly affected by kanamycin at $1mg{\cdot}L^{-1}$, but the number of shoots formed was significantly reduced at $2mg{\cdot}L^{-1}$, and no shoots were regenerated from any explants at $6mg{\cdot}L^{-1}$ or higher. Hypocotyl explants showed similar result as cotyledon. Kanamycin at $7mg{\cdot}L^{-1}$ may be adequate for selecting Chinese cabbage transformants. Hygromycin at $4mg{\cdot}L^{-1}$ or higher completely inhibited the growth and shoot regeneration of Chinese cabbage explants. Therefore, resistance gene to hygromycin may also be used as a selective marker for Chinese cabbage transformation. Carbenicillin and cefotaxime, the cephalosporin type of antibiotics, had little effect on shoot regeneration of Chinese cabbage explants. Since carbenicillin and cefotaxime have low toxicity to Chinese cabbage, they are suitable for use in tissue culture to eliminate Agrobacterium in transformation experiments after co-cultivation. Shoot regeneration from cotyledon and hypocotyl explants was significantly reduced in presence of $1mg{\cdot}L^{-1}$ phosphinotricin (PPT) and completely inhibited by $2mg{\cdot}L^{-1}$ or higher. PPT, same as antibiotics, may also be used to select transformed cells. Since Chinese cabbage is known to be recalcitrant to in vitro shoot regeneration compared to other Brassica species, even though lower levels of selectable markers result in more transformants but simultaneously allow more untransformed escapes to develop, lower levels of antibiotics and herbicides could be successfully used as a selectable marker to reduce selection pressure.

• Korean Journal of Breeding Science
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• v.49 no.3
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• pp.250-255
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• 2017

'Hwaweon 7' was developed from a cross between the African upland cultivar, 'Moroberekan' and 'Ilpumbyeo' based on marker-aided backcross selection. The recurrent parent, Ilpumbyeo is a high grain quality cultivar with medium to late maturity. Hwaweon 7 is nearly isogenic to Ilpumbyeo except a small Moroberekan introgressed segment on chromosome 6 harboring genes related with spikelets per panicle and internode diameter. The preliminary and replicated yield trial was conducted at Chungnam National University and Chungcheongnamdo Agricultural Research & Extension (CARES), Yesan in 2010 and 2011. The local adaptability test was carried out by the National Seed Management Office (NSMO) in 2012 and 2013. This cultivar was registered to NSMO with a cultivar designated as "Hwaweon7". This cultivar averaged 80cm in culm length and has a medium growth duration. Milled rice of "Hwaweon7" is translucent and the grain quality traits are comparable to those of the recurrent parent. The average yield potential of Hwaweon 7 in grain was about 6.48 MT/ha at the ordinary fertilizer level for two years about 5.1% higher than that of Ilpumbyeo due to increase in spikelets per panicle. This variety is tolerant to lodging and the tolerance is due to the APO1 gene controlling the base internode diameter on chromosome 6 introgressed from the donor parent, Moroberekan. The Apo1 gene would be useful in enhancing resistance to lodging in rice breeding program.

• Korean Journal of Breeding Science
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• v.50 no.4
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• pp.434-441
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• 2018

High-density genetic linkage mapping is critical for undertaking marker-assisted selection and confirming quantitative trait loci, as well as helping to build pseudomolecules of genomes. We constructed a genetic map using 94 $F_1$ populations generated from the interspecific cross between Korean cultivar "Wonwhang" (Pyrus pyrifolia, NCBI BioSample SAMN05196235) and European cultivar "Bartlett" (Pyrus communis). We designed a total of 24,267 SSR markers based on the genome sequences of "Wonwhang" for this. To select the markers that are linked to the traits important in pear breeding programs, SSR-containing genomic sequences were subjected to nucleotide sequence homology searches, which resulted in 510 SSR markers with high similarity to genes encoding proteins with putative functions such as transcription factors, resistance proteins, flowering time, and regulatory genes. Of these, 70 markers showed polymorphisms in parents and segregating populations and were used to construct a genetic linkage map, together with the unpublished 579 SNPs obtained from genotyping by sequencing analysis. The genetic linkage map covered 3,784.2 cM and the average distance between adjacent markers was 5.8 cM. Seventy SSR markers were distributed across 17 chromosomes with more than one locus.

• Journal of Acupuncture Research
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• v.31 no.2
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• pp.87-98
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• 2014

Objectives : We investigated whether snake venom toxin(SVT) from Vipera lebetina turanica sensitizes HT29 human epithelial colorectal cancer cells to tumor necrosis factor(TNF)-related apoptosis-inducing ligand(TRAIL) induced apoptosis in cancer cells. Methods : Cell viability assay was used to assess the inhibitory effect of TRAIL on cell growth of HT29 human colorectal cancer cells. And 6-diamidino-2-phenylindole(DAPI), terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay(TUNEL) staining assay were used to evaluate cell-apoptosis. Western blot analysis were conducted to observe apoptosis related proteins and death receptor. To assess whether the synergized inhibitory effect of SVT and TRAIL on reactive oxygen species(ROS) generation was reversed by strong anti-oxidative agent. Results : SVT with TRAIL inhibited HT29 cell growth different from TRAIL alone. Consistent with cell growth inhibition, the expression of TRAIL receptors; Expression of death receptor(DR)4 and DR5 was significantly increased and intrinsic pro-apoptotic cleaved caspase-3, -9 was subsequently increased together with increase of Bax/Bcl-2 ratio and extrinsic pro-apototic caspase-8 was also activated. In addition, the expression of anti-apoptotic survival proteins, a marker of TRAIL resistance(eg, cFLIP, survivin, X-linked inhibitor of apoptosis protein(XIAP) and Bcl-2) was suppressed by the combination treatment of SVT and TRAIL. Pretreatment with the ROS scavenger N-acetylcysteine abolished the SVT and TRAIL-induced upregulation of DR4 and DR5 expression and expression of the intrinsic pro-apoptotic caspase-3 and-9. Conclusion : The collective results suggest that SVT facilitates TRAIL-induced apoptosis in $HT_{29}$ human epithelial colorectal cancer cells through up-regulation of the TRAIL receptors; DR4 and DR5 and consecutive induction of bilateral apoptosis via regulating apoptosis related proteins.

• The Korea Journal of Herbology
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• v.20 no.4
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• pp.1-10
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• 2005

Objectives : We investigated the anti-aging effect on skin with the extract of the needles of red pine, Pinus densiflora. Methods : We measured various effects related to skin such as the anti-oxidant effect, the protection against ultraviolet (UV) irradiation, the inhibition of reactive oxygen species (ROS) generation, the induction of heat shock protein 70 (HSP70), the reduction of matrix metalloproteinase-2 (MMP-2) synthesis and senescent cell. Results : The results were as follows : The extract of the needles of red pine (RP) had the potent anti-oxidant effect and the ROS scavenging effect. Also RP preserved the systemic anti-oxidant enzyme system (superoxide dismutase and catalase) from UVB irradiation. RP protected the cell membrane from the damages induced by UVB irradiation. RP induced HSP70, a mediator of resistance to UVB irradiation. RP reduced the synthesis of MMP-2 induced by UVB irradiation. And RP inhibited the amount of senescent-associated (SA) ${\beta}-galactosidase$ staining, as a marker of replicative senescence. Conclusions : The results of our study indicate that the extract of the needles of red pine, Pinus densiflora, has anti-aging effects on skin.