• Title/Summary/Keyword: RD-PCR

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Supraclavicular BCG Lymphadenitis Noted at 21 Months after BCG Vaccination Confirmed by a Molecular Method (분자생물학적 방법으로 확진한 접종 21개월 후에 발생한 BCG 쇄골상부 림프절염 1례)

  • Lee, Min Hyun;Chae, Moon-Hee;Park, Kyoung Un;Cho, Hye-Kyung
    • Pediatric Infection and Vaccine
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    • v.21 no.2
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    • pp.139-143
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    • 2014
  • Bacille Calmette-$Gu\acute{e}rin$ (BCG) lymphadenitis is the most common complication of BCG vaccination. It commonly occurs in infants aged <6 months involving ipsilateral axillary lymph nodes. We described BCG lymphadenitis in a 22-month-old boy presenting swelling of left supraclavicular lymph node that was confirmed by real-time polymerase chain reaction (PCR) and the multiplex PCR targeting the region of difference (RD).

Expression of Mouse Adenosine Deaminase Gene in Transgenic Tobacco (Nicotiana tabacum L.) (형질전환 연초(Nicotiana tabacum L.)의 Mouse Adenosine Deaminase 유전자 발현)

  • 양덕춘;박지창;최광태;이정명
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.195-200
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    • 1995
  • The mammalian adenosine deaminase(ADA) gene was stably expressed in transgenic tobacco plane. The chimeric ADA gene 35S/35S/AMV/ADA/Tnos, has been constructed. This chimeric gene was introduced into the binary vector pRD400, which was thereafter mobilized into Agrobacterium tumefaiens strain MP90 harboring disarmed Ti-plasmid. The resulting strains were used to transform Nicofiana tabacum L. using the leaf disc. Incorporation of the chimeric gene into plant were confirmed by PCR and Northern blot analyses. Immunoblot analysis showed that ADA protein was successfully synthesized in the transgenic tobacco plants.

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The First Identified Citrus tristeza virus Isolate of Turkey Contains a Mixture of Mild and Severe Strains

  • Cevik, Bayram;Yardimci, Nejla;Korkmaz, Sava
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.31-41
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    • 2013
  • The presence of Citrus tristeza virus (CTV) has previously been reported in citrus growing regions of Turkey. All serologically and biologically characterized isolates including I$\breve{g}$d${\i}$r, which was the first identified CTV isolates from Turkey, were considered mild isolates. In this study, molecular characteristics of the I d r isolate were determined by different methods. Analysis of the I$\breve{g}$d${\i}$r isolate by western blot and BD-RT-PCR assays showed the presence of MCA13 epitope, predominantly found in severe isolates, in the I$\breve{g}$d${\i}$r isolate revealing that it contains a severe component. For further characterization, the coat protein (CP) and the RNA-depen-dent RNA polymerase (RdRp) genes representing the 3' and 5' half of CTV genome, respectively, were amplified from dsRNA by RT-PCR. Both genes were cloned separately and two clones for each gene were sequenced. Comparisons of nucleotide and deduced amino acid sequences showed that while two CP gene sequences were identical, two RdRp clones showed only 90% and 91% sequence identity in their nucleotide and amino acid sequences, respectively, suggesting a mixed infection with different strains. Phylogenetic analyses of the CP and RdRp genes of I$\breve{g}$d${\i}$r isolate with previously characterized CTV isolates from different citrus growing regions showed that the CP gene was clustered with NZRB-TH30, a resistance breaking isolate from New Zealand, clearly showing the presence of severe component. Furthermore, two different clones of the RdRp gene were clustered separately with different CTV isolates with a diverse biological activity. While the RdRp-1 was clustered with T30 and T385, two well-characterized mild isolates from Florida and Spain, respectively, the RdRp-2 was most closely related to NZRB-G90 and NZRB-TH30, two well-characterized resistance breaking and stem pitting (SP) isolates from New Zealand confirming the mixed infection. These results clearly demonstrated that the I$\breve{g}$d${\i}$r isolate, which was previously described as biologically a mild isolate, actually contains a mixture of mild and severe strains.

Differential Expression of Osteonectin in the Rat Developing Molars

  • Kim, Jung-Ha;Yoo, Hong-Il;Oh, Min-Hee;Yang, So-Young;Kim, Min-Seok;Kim, Sun-Hun
    • International Journal of Oral Biology
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    • v.37 no.2
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    • pp.51-56
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    • 2012
  • Tooth development involves bud, cap, bell and hard tissue formation stages, each of which is tightly controlled by regulatory molecules. The aim of this study was to identify genes that are differentially expressed during dental hard tissue differentiation. Sprague-Dawley rats at postnatal days 3, 6 and 9 were used in the analysis. Differential display RT-PCR (DD-PCR) was used to screen differentially expressed genes between the 2nd (root formation stage, during mineralization) and 3rd (cap stage, before mineralization) molar germs at postnatal day 9. The DNA detected in the 2nd molar germs showed homology to osteonectin only (GenBank accession no. NM_012656.1). The level of osteonectin mRNA expression was much higher in the 2nd molar germs than in the 3rd molar germs and was found to increase in a time-dependent manner from the early bell stage to the root formation stage in the 2nd molar germs. The pattern of osteonectin protein expression was consistent with these RT-PCR results. Osteonectin protein was found by immunofluorescent analysis to localize in odontoblasts and preodontoblasts rather than the dentin matrix itself. Further studies are needed to validate the involvement of osteonectin in mineralization and root formation.

Study on the Market Conditions and Quality Evaluation Methods of post-consumer Recycled Polyethylene Terephthalate (PCR PET) Flake (재생 페트 플레이크의 시장 현황과 품질 평가 방안 연구)

  • Joo, Minjung;Suh, Sang Uk;Lee, Ka Eun;Oh, Jae Young
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.26 no.1
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    • pp.41-46
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    • 2020
  • PCR PET flake's market endeavors difficult situations of its oversupply and decreasing demands in South Korea. Since China banned the import of most recycled plastics, flake producers who mostly export the flakes lost their biggest market. The producers struggled to survive in the competitions with PCR PET flake from EU and 3rd countries but it was challenging due to substandard quality and increasing cost. Attempts to improve the quality of PCR PET flake have been made but they were only an individual company's efforts. The objective of this study was to understand the market status on PCR PET flake in South Korea and to present suggestions for improving its quality. The results of the questionnaires targeted to flake producers showed that no testing methods on PCR PET flake were standardized and there were critical factors to the quality such as moisture content, contaminants, and viscosity. Case studies of US, Japan and other countries had been done especially about testing methods and 76 samples from 21 companies were tested according to those methods. Based on the results, the final factors were decided as contaminants, moisture content, alkalinity index and intrinsic viscosity. There is a plan to standardize testing methods and they could be guidelines to improve the business competitiveness of PCR PET flake in South Korea.

Effects of Sagunjatang and Sagunja-tang plus Mylabris phalerata on Human Stomach Cancer Cells (사군자탕(四君子湯) 및 사군자탕가반묘(四君子湯加斑猫)가 위암세포에 미치는 영향)

  • Jung, Woo-Young;Ryu, Bong-Ha;Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Ki-Won;Sin, Hyeon-Su
    • The Journal of Internal Korean Medicine
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    • v.22 no.4
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    • pp.579-587
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    • 2001
  • The efficacy of Sagunja-tang and Sagunja-tang plus Mylabris phalerata against the human stomach cancer was examined and molecular biological fight of its actions was studied. In the efficacy test of anti-stomach cancer cells growth using the MTT assay, administration of Sagunja-tang resulted in no significant change of stomach cancer cells growth, with the control group. Administration of Sagunja-tang plus Mylabris phalerata resulted in a decrease of stomach cancer cells growth in proportion to the concentration of mylabris phalerata and time, which was significantly different from the control group(significance recognized when p<0.05). In the test using the apoptosis assay, administration of Sagunja-tang showed an increase in apoptosis of human stomach cancer cells, with no significant difference from the control group. Administrating Sagunja-tang plus Mylabris phalerata showed an increase in apoptosis of stomach cancer cells in proportion to the concentration of mylabris phalerata and time, which was significantly different from the control group(significance recognized when p<0.05). In the test using the quantitative RT-PCR to examine stomach cancer cells growth and revelation of apoptosis related genes, administrating Sagunja-tang plus Mylabris phalerata resulted in a decrease of Bcl-2, an anti-apoptosis gene, in proportiong to concentration. No significant change was examined in the revelation of CDK1, Cdc2, Cyclin D1, PCNA, c-myc, which are genes related to the stomach cancer cells growth, and Bax, Bel-XL, the genes related to apoptosis, and p53. Referring to the results above, Sagunja-tang plus Mylabris phalerata may be considered to have an anti-growth efficacy against human stomach cancer cells, and an inducement efficacy. Therefore, it can be clinically implemented in the human stomach cancer.

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Nucleotide Sequence Analysis of the RNA-dependent RNA Polymerase Gene of Infectious Pancreatic Necrosis Virus DRT Strain

  • Lee, Hyung-Hoan;Chung, Hye-Kyung;Lee, Seong-Hun
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.264-269
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    • 1994
  • To determine the nucleotide sequence of the ds RNA segment B containing the RNA dependent RNA polymerase (RdRp) gene of the DRT strain of infectious pancreatic necrosis virus (lPNV), the cDNA of the ds RNA segment B of the DRT strain of IPNV was synthesized using the reverse transcriptase (RT)-polymerase chain reaction (PCR) and its cDNA nucleotide sequence was determined. The DRT segment B was 2, 783 bp long and contained only a single long open reading frame (ORF) of 2, 535 bp in length. This ORF nucleotides encoded the VPl protein, the putative RdRp of IPNV. The VPl protein comsisted of 845 amino acids. The molecular weight of the RdRp, as deduced from the nucleotide sequence, is 94, 426. The nucleotide sequence of the ORF of the DRT showed 89.7% homology to the Jasper strain, but 80.8% to the Sp strain. The amino acid sequence of the ORF of the DRT sho.wed 97.6% homology to the Jasper strain, but 88.7% to the Sp strain. The conserved GTP-binding motif was detected in VPl protein.

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Identification of a norovirus from diarrheic dog in Gwangju, Republic of Korea

  • Ba-Ra-Da Koh;Su-Yeon Seo;Ga-Hoi Choi;Byeong-Cheol Yoon
    • Korean Journal of Veterinary Service
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    • v.46 no.3
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    • pp.235-241
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    • 2023
  • Noroviruses are a major cause of gastroenteritis in humans and animals worldwide. In 2021, canine norovirus (CNoV) infection was detected at an animal clinic in Gwangju area, South Korea. A semi-nested polymerase chain reaction was developed to amplify a 478 bp fragment of the RdRp gene of CNoV. The phylogenetic analysis of this fragment confirmed the strain to be genogroup IV.2 (Dog/GIV.2/gw/s377/2021/KOR), which exhibited the highest similarity to the feline NoV strain GIV.2/CU081210E/USA/2010 (accession no. NC_045762) with 95.1% nucleotide (nt) identity and 98.7% amino acid (aa) identity. These research findings indicate that the detected norovirus in dogs is genetically similar to a feline-origin norovirus, suggesting easy cross-species transmission among animals.

Characterization of Odontoblasts in Supernumerary Tooth-derived Dental Pulp Stem Cells between Passages by Real-Time PCR (과잉치 치수유래 줄기세포의 Real-time PCR에 의한 계대간 상아질모세포 발현 특성)

  • Ji, Sangeun;Song, Sol;Lee, Joonhaeng;Kim, Jongbin;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.48 no.3
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    • pp.291-301
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    • 2021
  • The aim of this study is to compare the properties of odontoblast gene of early passage cells and late passage cells derived from impacted maxillary supernumerary teeth. Impacted supernumerary teeth with maxilla were extracted from 12 patients (8 males, 4 females) between 6 - 9 years old without medical history. Real-time polymerase chain reaction (PCR) was conducted to compare characterization of odontoblast cell in the 3rd and 10th passage, and between with bone inducing additive group and without additive group. Genes for odontoblasts characteristics are osteonectin (ONT), alkaline phosphatase (ALP), osteocalcin (OCN), dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP). The level of gene expression was in a decreasing order of ONT, ALP, OCN, DMP-1 and DSPP in the 3rd passage, and in decreasing order of ONT, DMP-1, OCN, ALP, and DSPP in the 10th passage in the undifferentiation and differentiation group. The order of ONT, DMP-1, and OCN did not changed. ALP and DMP-1 were switched in order. ALP and DMP-1 may be used as important markers for differentiating between the 3rd passage and 10th passage cells. Considering that supernumerary tooth was extracted young age and the time required to cultured 10th passage was short, supernumerary tooth can be considered a useful donor site of dental pulp stem cells.

Development of Narrow Leaf Type Cultivar "Konhee" in Zoysiagrass (세엽 한국잔디류 신품종 '건희(Konhee)' 육성)

  • 김두환;이재필;김종빈;모숙연
    • Asian Journal of Turfgrass Science
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    • v.13 no.3
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    • pp.147-151
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    • 1999
  • "Konhee" zoysiagrass[Zoysia matrella (L) Merr.](Patent registration no. 2000-723), a vegetative cultivar, was developed by the Dept. of Horticultural Science, Konkuk University, Seoul. "Konhee" was selected from the cross, ZKV6$\times$ZKV10, in 1997 and F1 seed were produced in the greenhouse in 1996. "Konhee" is superior to the other fine leaf zoysiagrass lines in many traits such as erect type, plant height($8.5\pm$2.0cm), short 3rd stolon length($3.4\pm$0.5cm), dark green, fine leaf($2.3\pm$0.2mm), low first sheath height($0.9\pm$0.2cm), rapid establishment and recoverage, many stolon number, and high shoot density. When "Konhee" was compared to the five other zoysisagrass lines at the DNA level using 35 PCR primers, it had the specific bands with primer No. 740, 544, 765, 772 by RAPD analysis.No. 740, 544, 765, 772 by RAPD analysis.

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