• Title/Summary/Keyword: RPO

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Chloroplast Genome Evolution in Early Diverged Leptosporangiate Ferns

  • Kim, Hyoung Tae;Chung, Myong Gi;Kim, Ki-Joong
    • Molecules and Cells
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    • v.37 no.5
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    • pp.372-382
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    • 2014
  • In this study, the chloroplast (cp) genome sequences from three early diverged leptosporangiate ferns were completed and analyzed in order to understand the evolution of the genome of the fern lineages. The complete cp genome sequence of Osmunda cinnamomea (Osmundales) was 142,812 base pairs (bp). The cp genome structure was similar to that of eusporangiate ferns. The gene/intron losses that frequently occurred in the cp genome of leptosporangiate ferns were not found in the cp genome of O. cinnamomea. In addition, putative RNA editing sites in the cp genome were rare in O. cinnamomea, even though the sites were frequently predicted to be present in leptosporangiate ferns. The complete cp genome sequence of Diplopterygium glaucum (Gleicheniales) was 151,007 bp and has a 9.7 kb inversion between the trnL-CAA and trnV-GCA genes when compared to O. cinnamomea. Several repeated sequences were detected around the inversion break points. The complete cp genome sequence of Lygodium japonicum (Schizaeales) was 157,142 bp and a deletion of the rpoC1 intron was detected. This intron loss was shared by all of the studied species of the genus Lygodium. The GC contents and the effective numbers of codons (ENCs) in ferns varied significantly when compared to seed plants. The ENC values of the early diverged leptosporangiate ferns showed intermediate levels between eusporangiate and core leptosporangiate ferns. However, our phylogenetic tree based on all of the cp gene sequences clearly indicated that the cp genome similarity between O. cinnamomea (Osmundales) and eusporangiate ferns are symplesiomorphies, rather than synapomorphies. Therefore, our data is in agreement with the view that Osmundales is a distinct early diverged lineage in the leptosporangiate ferns.

The first case report on Vibrio atlanticus infection in cultured tiger puffer, Takifugu rubripes (우리나라 양식 자주복, Takifugu rubripes (Temminck & Schlegel, 1850)에서 발생한 Vibrio atlanticus의 첫 감염 사례 보고)

  • Lee, Nam-Sil;Cho, Miyoung;Jung, Sung-hee;Won, Kyoungmi
    • Journal of fish pathology
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    • v.31 no.2
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    • pp.87-92
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    • 2018
  • A case of disease was occurred in tiger puffer, Takifugu rubripes, reared in circulated system (water temperature was about $20^{\circ}C$) on land in Korea. Diseased fish showed a unique external sign, the cloudy skin ulcer in various sizes. We investigated the cause of disease, and isolated one pure cultured bacterium, TP01. This had 99.7% homology with Vibrio atlanticus by sequence analysis of the 16S rRNA and rpoA genes. TP01 showed some differences with other reported V. atlanticus strains in the biochemical characteristics as like Voges-Proskauer test and gelatin hydrolysis.

Development of Oligonucleotide Chip for Detection of Drug-Resistant Mycobacterium Tuberculosis (약제내성 결핵균의 검출을 위한 Oligonucleotide Chip의 개발)

  • Song, Eunsil;Park, Heekyung;Jang, Hyunjung;Kim, Hyomyung;Chang, Chulhun L.;Kim, Cheolmin
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.1
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    • pp.41-58
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    • 2003
  • Background : The resurgence of tuberculosis and the widespread emergence of multidrug-resistant M. tuberculosis have emphasized the importance of rapid and accurate diagnostic procedures. Recently, the oligonucleotide chip has proven to be a useful tool in the rapid diagnosis of infectious diseases. The purpose of this study was to rapidly and accurately detect specific mutations in the rpoB, katG and rpsL genes associated with rifampin, isoniazid and streptomycin resistance in M. tuberculosis, respectively, using a single oligonucleotide chip. Method : For detection of drug-resistance, 7 wild-type and 13 mutant-type probes for rifampin, 2 wild-type and 3 mutant-type probes for isoniazid, and 2 wild-type and 2 mutant-type probes for streptomycin were designed and spotted onto glass slides. Fifty-five cultured samples of M. tuberculosis were amplified by PCR, and then underwent hybridization and scanning. Direct sequencing was done to verify the results from the oligonucleotide chip and to analyze the types of mutations. Result : Thirty-five cases out of 40 rifampin-resistant strains(~88%) had mutations in the rpoB gene. One case had a new mutation(D516F, GAC R TTC) and another known mutation together. Twenty cases out of 42 isoniazid-resistant strains(~50%) had mutations in the katG gene, while 7 cases out of 9 streptomycin-resistant strains(~78%) had mutations in the rpsL gene. From these results, the oligonucleotide chip was confirmed to be able to detect the most frequent mutations from the genes associated with rifampin, isoniazid and streptomycin resistance. The results proved that the drug-resistance detection probes were specific. When the results from the oligonucleotide chip and DNA sequencing were compared, the types of mutations were exactly matched. Conclusion : The diagnostic oligonucleotide chip with mutation specific probes for drug resistance is a very reliable and useful tool for the rapid and accurate diagnosis of drug resistance against rifampin, isoniazid and streptomycin in M. tuberculosis infections.

Application for Identification of Food Raw Materials by PCR using Universal Primer (일반 프라이머를 이용한 PCR의 식품원료 진위 판별에 적용)

  • Park, Yong-Chjun;Jin, Sang-Ook;Lim, Ji-Young;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Han, Sang-Bae;Lee, Sang-Jae;Lee, Kwang-Ho;Yoon, Hae-Seong
    • Journal of Food Hygiene and Safety
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    • v.27 no.3
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    • pp.317-324
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    • 2012
  • In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

Clinical Meaning of INNO-LiPA Test in the Diagnosis of Rifampin Resistant Tuberculosis (Rifampin 내성 결핵의 진단에서 INNO-LiPA 검사법의 임상적 의미)

  • Chang, Yoon Soo;Kim, Young;Lee, Chang Youl;Choi, Jong Rak;Kim, Hyung Jung;Ahn, Chul Min;Kim, Sung Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.4
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    • pp.344-352
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    • 2003
  • Background : The prevalence of multidrug resistant tuberculosis (MDR-TB), resistant to isoniazid (INH) and rifampin (RFP), was 5.3% worldwide in 1995 and its increment has raised important public health problems. Resistance to RFP, one of the key drugs in the treatment of tuberculosis, results in grim clinical outcome. Recently rapid detection of RFP-resistant mutations in rpoB gene based on PCR method has become available. This study evaluated the prevalence of RFP resistance in first diagnosed, treatment failure, and recurred patients using INNO-LiPA test, and compared the results of INNO-LiPA with those of conventional mycobacterial drug susceptibility test. Methods : Forty-six patients, who were diagnosed of pulmonary tuberculosis and had revealed positive sputum AFB smear, were enrolled in this study from 1998 to 2002. The cases were classified as one three groups; first diagnosed, treatment failure, or recurred. RFP resistance was studied using an INNO-LiPA Rif. TB kit and compared with that obtained from drug susceptibility based on M. tuberculosis culture study. Results : Twenty-one out of 46 patients were enrolled under first diagnosis of pulmonary tuberculosis, 17 under treatment failure with first line drugs, and 8 under recurrence. The positive and negative predictive values of INNO-LiPA test in diagnosis in RFP resistant tuberculosis compared with conventional mycobacterial drug susceptibility test were 85.7% and 76.0%, respectively. INNO-LiPA result revealed rpoB gene mutation in 20 (80.0%) out of 25 patients who were diagnosed as treatment failure or recurrence, but in only 4 (19.0%) out of 21 patients who were first diagnosed as pulmonary tuberculosis. Conclusion : This study showed that RFP resistance could be diagnosed rapidly and accurately using INNO-LiPA test and that this test might be helpful for choosing second line anti-mycobacterial drugs. It might be of great help in clinical diagnosis and decision when used in complimentarily with drug susceptibility test based on M. tuberculosis culture.

Detection Method for Identification of Pueraria mirifica (Thai kudzu) in Processed Foods (가공식품 중 태국칡(Pueraria mirifica) 혼입 판별법 개발)

  • Park, Yong-Chjun;Jin, Sang-Wook;Kim, Mi-Ra;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.27 no.4
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    • pp.466-472
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    • 2012
  • In this study, ribulose bisphosphate carboxylase (rbcL), RNApolymeraseC (rpoC1), intergenic spacer (psbA-trnH), and second internal transcribed spacer (ITS2) as identification markers for discrimination of P. mirifica in foods were selected. To be primer design, we obtained 719 bp, 520 bp, 348 bp, and 507 bp amplicon using universal primers from selected regions of P. mirifica. The regions of rbcL, rpoC1, and psbA-trnH were not proper for design primers because of high homology about P. mirifica, P. lobata, and B. superba. But, we had designed 4 pairs of oligonucleotide primers from ITS2 gene. Predicted amplicon from P. mirifica were obtained 137 bp and 216 bp using finally designed primers SFI12-miri-6F/SFI12-miri-7R and SFI12-miri-6F/SFI12-miri-8R, respectively. The species-specific primers distinguished P. mirifica from related species were able to apply food materials and processed foods. The developed PCR method would be applicable to food safety management for illegally distributed products in markets and internet shopping malls.

High-efficient Expression of Porcine IL-2 with Recombinant Baculovirus Infected Silkworm, Bombyx mori

  • Inumaru, Shigeki;KokuHo, Takehiro;Yada, Takashi;Kiuchi, Makoto;Miyazawa, Mitsusuhiro
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.146-149
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    • 2000
  • Biologically active porcine Interleukin-2(poIL-2) was produced from in vitro and in viva baculovirus expression system, namely the Autographa californica nuclear polyhedrosis virus (ACNPV)-cell culture system and the Hybrid nucler polthedrosis virus (HyNPV)-sillkworm larva system. The concentration of the recombinant poIL-2(rpoIL-2) in the larvae hemolymph was 1 to 3 mg/mL, which was about 7 to 20 times those of the cell culture systems. The level of this expression efficiency is equal to that with transgenic livestock, secretion products in milk.

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Nontuberculous Mycobacterial Lung Disease Caused by Mycobacterium terrae in a Patient with Bronchiectasis

  • Koh, Won-Jung;Choi, Go-Eun;Lee, Nam-Yong;Shin, Sung-Jae
    • Tuberculosis and Respiratory Diseases
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    • v.72 no.2
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    • pp.173-176
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    • 2012
  • We report a rare case of lung disease caused by Mycobacterium terrae in a previously healthy woman. A 45-year-old woman was referred to our hospital due to a chronic cough with sputum. A computed tomography scan of the chest revealed bronchiolitis in conjuction with bronchiectasis in both lungs. Nontuberculous mycobacteria were identified and isolated from the bronchoalveolar lavage fluid collected from each lung. All isolates were identified as M. terrae by various molecular methods that characterized the rpoB and hsp65 gene sequences. Antibiotic therapy using clarithromycin, rifampin, and ethambutol improved the patient's condition and successfully resulted in sputum conversion.

Preliminary search of intraspecific chloroplast DNA variation of nine evergreen broad leaved plants in East Asia

  • Lee, Jung-Hyun;Lee, Byoung-Yoon;Choi, Byoung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.41 no.3
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    • pp.194-201
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    • 2011
  • In order to acquire information on chloroplast DNA markers to evaluate the genetic diversity of evergreen broad leaved plants, we investigated the intraspecific variation of cpDNA in eight non-coding regions of nine species commonly distributed in East Asia. Although no variations were detected in psbA-trnH, rpoB-trnC, rpl16 and atpB-rbcL regions, a relatively large amount of intraspecific variations was detected in the psbC-trnS, rps16 and trnL-F regions. These results suggested that these three cpDNA markers are suitable to assess genetic diversity of the species investigated in this study. In contrast, intraspecific variations were detected in seven taxa except Hedera rhombea and Neolitsea aciculata. Neolitsea sericea and the taxa of Quercus had many polymorphic sites.

Reclassification of a Carboxydobacterium, Acinetobacter sp. Strain JC1 DSM3803, as Mycobacterium sp. Strain JC1 DSM 3803

  • Taeksun Song;Lee, Hyeyoung;Park, Yong-Ha;Kim, Eungbin;Ro, Young-Tae;Kim, Si-Wouk;Kim, Young-Min
    • Journal of Microbiology
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    • v.40 no.3
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    • pp.237-240
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    • 2002
  • A carboxydotrophic bacterium, isolated from a soil sample in Seoul, was classified initially as Acinetobacter sp. strain JC1 DSM 3803. Chemotaxanomic properties, analysis of the 16s rDNA sequence, fatty acid content, and molecular Phylogenetic analysis based on rpoB gene, however, suggested that this bacterium belongs to the genus, Mycobacterium. On the basis of this evidence, it is proposed that Acinetobacter sp. strain JC1 DSM 3803 be reclassified as Mycobacterium sp. strain JC1 DSM 3803.