• Title/Summary/Keyword: rpoD

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Regulation of stf Operon Expression and Its Virulence (살모넬라가 발현하는 stf 오페론의 조절과 병원성 인자로서의 기능)

  • Kim Sam-Woong;Kim Young-Hee;Kang Ho-Young
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.553-560
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    • 2005
  • The stf (Salmonella typhimurium fimbriae) operon consisting of stfA(CDEFG assumes to encode putative fimbriae. The complete stf operon is existed in S. typhimurium and S. choleraesuis, whereas it is absent in S. typhi. Analyses of the amino acid residues between major subunit StfA of the Stf fimbriae and those of known other fimbriaes suggested that Stf belongs to class I type fimbriae. Through comparison of StfD chaperone with the other fimbrial chaperones, and of C-terminus in subunits of Stf fimbriae, it belongs to FGS (with a short Fl-G1 loop) subfamily. In order to investigate the expression of stf operon, we have constructed a Salmonella strain containing a chromosomal stfA::lacZYA transcriptional fusion, resulting in S. typhimurium $_X8532$. The strain $_X8532$ lacked the expression of \beta-galactosidase$ under normal culture conditions. However, with longer incubation time of the S. typhimurium $_X8532$, we have isolated 21 individual strains exhibiting $Lac^+$ phenotype. $Lac^+$ phenotype was appeared as approximately 0.03 frequency per generation. All isolates expressed lacZ constitutively in the various environmental conditions. Various global regulatory proteins including RpoS, OmpR, and CpxR were not involved in the regulation of the stf operon. A S. typhimurium $_X8661$ mutant lacking stfAC function attenuated 6.7 folds more than that of wild type $_X3761$ in the mouse virulence test, suggesting in the somehow involved in the Salmonella pathogenesis.

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.

First Report of Bacterial Spot Disease Caused by Pseudomonas capsici on Castor Bean in Korea (Pseudomonas capsici에 의한 아주까리 세균점무늬병의 국내 첫 보고)

  • Heeil Do;Seung Yeup Lee;Bang Wool Lee;Hyeonheui Ham;Mi-Hyun Lee;Young Kee Lee
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.440-444
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    • 2023
  • In August 2021, water-soaking symptoms of bacterial spot disease were observed on castor bean in a field in Gangseo District, Busan. Bacteria isolated from the lesion when cultured on tryptic soy agar appeared to be nonmucoid and pale green. To confirm whether the isolates were the causative agent of the spot disease, they were inoculated onto healthy castor bean plants. The same symptoms were observed on the inoculated tissue, and the bacteria were reisolated from the lesion. Furthermore, the isolates were consistent with the biochemical and physiological features of Pseudomonas capsici. Sequencing analysis using 16S rRNA and housekeeping genes (gyrB, rpoD) showed that the isolates shared a high sequence similarity with P. capsici. These results confirmed that the strains belonged to P. capsici. To our knowledge, this is the first report of bacterial spot disease caused by P. capsici on castor bean in Korea.

Plastome Phylogenomics of Commelinaceae Mirb. (Commelinales): Insights into Genome Evolution and Phylogenetic Relationships

  • Joonhyung Jung;Joo-Hwan Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.69-69
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    • 2022
  • Commelinaceae (Commelinales), consist of three subfamiles and 40 genera, are distributed in the Old and New world, except Europe. This family is commonly known as dayflower and spiderwort due to their short bloom time and a viscous stem secretion. Although, several morphological and molecular analysis were conducted, the relationships among the genera are still ambiguous. The rapid advances in next-generation sequencing (NGS) enable us to do genomic research widely. Here, we assembled 12 new plastomes of Commelinaceae including Cartonematoideae and compared with previously published data. We identified pseudogened accD and rpoA in Commelinoideae taxa. Phylogenetic analysis inferred from 78 protein-coding genes showed that Rhopalephora scaberrima was nested within Aneilema. Also, there is a need to revise the subtribal relationships in Tradescantieae. This study will contribute to define the genome structures, phylogenetic and biogeographic studies of Commelinaceae.

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Occurrence of Bacterial Stem Rot of Ranunculus asiaticus Caused by Pseudomonas marginalis in Korea

  • Li, Weilan;Ten, Leonid N.;Kim, Seung-Han;Lee, Seung-Yeol;Jung, Hee-Young
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.138-144
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    • 2018
  • In December 2016, stem rot symptoms were observed on Persian buttercup (Ranunculus asiaticus) plants in Chilgok, Gyeongbuk, Korea. In the early stage of the disease, several black spots appeared on the stem of infected plants. As the disease progressed, the infected stem cleaved and wilted. The causal agent was isolated from a lesion and incubated on Reasoner's 2A (R2A) agar at $25^{\circ}C$. Total genomic DNA was extracted for phylogenetic analysis. Based on the 16S rRNA gene analysis, the isolated strain was found to belong to the genus Pseudomonas. To identify the isolated bacterial strain at the species level, the nucleotide sequences of the gyrase B (gyrB) and RNA polymerase D (rpoD) genes were obtained and compared with the sequences in the GenBank database. As the result, the causal agent of the stem rot disease was identified as Pseudomonas marginalis. To determine the pathogenicity of the isolated bacterial strain, it was inoculated into the stem of healthy R. asiaticus plant, the inoculated plant showed a lesion with the same characteristics as the naturally infected plant. Based on these results, this is the first report of bacterial stem rot on R. asiaticus caused by P. marginalis in Korea.

Distribution of Bacterial Angular Leaf Spot of Strawberry and Characterization of Xanthomonas fragariae Strains from Korea (한국의 딸기세균모무늬병 발생분포 및 딸기세균모무늬병균 특성조사)

  • Yoon, Myung-Ju;Myung, Inn-Shik;Lee, Jae-Yeon;Kim, You-Shin;Lee, Yong-Hwan;Kim, Dae-Young;Lee, Young-Ki
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.9-17
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    • 2016
  • Nationwide survey for angular leaf spot (ALS) of strawberry caused by Xanthomonas fragariae, a quarantine disease in Korea, was performed in November 2012. In the survey, ALS was observed in eighty eight farmers' fields of Sukok, Jinju and Okjong, Hadong in Gyeongnam Province, and one field in Namwon of Jeollabuk Province. The infected field of Namwon closed immediately after the disease diagnosed ALS. In detailed survey of Sukok and Okjong areas during February 2012 to January 2015, ALS occurrence decreased from 45% farmer's fields on December 2012 to 5% on January 2015, and from 38% on November 2013 to 5% on January 2015, respectively. Phenotypic characteristics of the Korean strains were similar to those of the type strain of X. fragariae. A multilocus sequence analysis of Korean strains of X. fragariae was conducted using four genes; dnaK, fyuA, gyrB, and rpoD. All the Korean strains had the same sequences of the four genes. The concatenated sequences of the Korean strains shared 100% with that of the type strain of X. fragariae. All strawberry cultivars tested were susceptible to the strains of X. fragariae two weeks after inoculation. The inoculated sites were necrosis and expanded, which were rated 4 based on evaluation of inoculation site.

Bacterial Spot Disease of Green Pumpkin by Pseudomonas syringae pv. syringae (Pseudomonas syringae pv. syringae에 의한 애호박 세균점무늬병)

  • Park, Kyoung-Soo;Kim, Young-Tak;Kim, Hye-Seong;Lee, Ji-Hye;Lee, Hyok-In;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.22 no.3
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    • pp.158-167
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    • 2016
  • A pathogen that causes a new disease on green pumpkin in the nursery and the field was characterized and identified. Symptoms of the disease on green pumpkin were water soaking lesions and spots with strong yellow halo on leaf, brown lesion on flower, and yellow spot on fruit. The bacterial isolates from the leaf spot were pathogenic on the 8 curcubitaceae crop plants, green pumpkin, figleaf gourd, wax gourd, young pumpkin, zucchini, cucumber, melon, and oriental melon, whereas they did not cause the disease on sweet pumpkin and watermelon. They were Gram-negative, rod shape with polar flagella, fluorescent on King's B agar and LOPAT group 1a by LOPAT test. Their Biolog substrate utilization patterns were similar to Pseudomonas syringae pv. syringae's in Biolog database. Phylogenetic trees with 16S rRNA gene sequences and multilocus sequence typing (MLST) with nucleotide sequences of 4 housekeeping genes, gapA, gltA, gyrB, rpoD and those of P. syringae complex strains in the Plant Associated and Environmental Microbes Database (PAMDB) showed that the green pumpkin isolates formed in the same clade with P. syringae pv. syringae strains. The clade in MLST tree was in the genomospecies 1 group. The phenotypic and genotypic characteristics suggested that the isolates from green pumpkin lesion were P. syringae pv. syringae.

Occurrence of Leaf Spot Disease on Watermelon Caused by Pseudomonas syringae pv. syringae (Pseudomonas syringae pv. syringae에 의한 수박 잎점무늬병의 발생)

  • Park, Kyoung-Soo;Lee, Ji-Hye;Kim, Young-Tak;Kim, Hye-Seong;Lee, June-woo;Lee, Hyun-Su;Lee, Hyok-In;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.27 no.4
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    • pp.180-186
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    • 2021
  • Typical bacterial symptoms, water-soaking brown and black leaf spots with yellow halo, were observed on watermelon seedlings in nursery and field of Gyeongnam and Jeonnam provinces. Bacterial isolates from the lesion showed strong pathogenicity on watermelon and zucchini. One of them was rod-shaped with 4 polar flagella by observation of transmission electron microscopy. They belonged to LOPAT group 1. The phylogenical trees with nucleotide sequences of 16S rRNA and multi-locus sequencing typing with the 4 house-keeping genes (gapA, gltA, gyrB, and rpoD) of the isolates showed they were highly homologous to Pseudomonas syringae pv. syringae and grouped together with them, indicating that they were appeared as P. syringae genomospecies group 1. Morphological, physiological, and genetical characteristics of the isolates suggested they are P. syringae pv. syringae. We believe this is the first report that P. syringae pv. syringae caused leaf spot disease on watermelon in the Republic of Korea.

Effect of Carbon Couch Side Rail and Vac-lok In case of Lung RPO irradiation (Lung RPO 선량전달시, Carbon Couch Side Rail과 Vac-lok이 미치는 영향)

  • Kim, Seok Min;Gwak, Geun Tak;Lee, Seung Hun;Kim, Jung Soo;Kwon, Hyoung Cheol;Kim, Yang Su;Lee, Sun Young
    • The Journal of Korean Society for Radiation Therapy
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    • v.30 no.1_2
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    • pp.27-34
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    • 2018
  • Purpose : To evaluate the effect of carbon couch side rail and vacuum immobilization device in case of lung RPO irradiation. Materials and Methods : The 10, 20, 30 mm thickness of vac-lok's right side were obtained. To measure of doses, glass dosimeters were used and measured reference point is left lung center at the phantom. A, B, C, and D points are left, right, down, and up directions based on the center point. In the state of Side-Rail-Out, place the without vac-lok, with the thickness of 10, 20, and 30 mm vac-lok. After the glass dosimeters was inserted in center, A, B, C, and D points, 100 MU of 6 MV X-ray were irradiated to the referenced center point in the condition of $10{\times}10cm^2$ field size, SAD 100 cm, gantry angle 225, 300 MU/min dose rate. Five measurements were made for each point. In the state of Side-Rail-In, five measurement were made for each point under the same conditions. The average is measured on each of the five Side-Rail-Out and Side-Rail-In measurements. Results : In the presence of side rail, the dose reduction ratio was -11.8 %, -12.3 %, -4.1 %, -12.3 %, -7.3 % for each A, B, C, and D points. In the state of Side-Rail-Out, the dose reduction ratio for the using 10 mm thickness of vac-lok was -0.9 % than without vac-lok. The dose reduction ratio for the using 20 mm thickness of vac-lok was -2.0 %, for the using 30 mm thickness of the vac-lok was -3.0 % than without vac-lok. In the state of Side-Rail-In, the dose reduction ratio for the using 10 mm thickness of vac-lok was -1.0 % than without vac-lok. The dose reduction ratio for the using 20 mm vac-lok was -2.1 %, for the using 30 mm vac-lok was -3.0 % than without vac-lok. Based on the value of no vac-lok dose in the Side-Rail-In state, The dose reduction ratios for the using 10 mm, 20 mm and 30 mm thickness of vac-loks In the Side-Rail-Out that the center point were -12.7 %, -13.7 %, -14.2 % and -12.8 %, -13.8 %, -14.5 % respectively at point A. The dose reduction ratios for the same conditions to the B point were -4.9 %, -6.1 %, -7.1 % and -13.4 %, -14.4 %, -15.5 % respectively at point C. The dose reduction ratios for the same conditions to the D point were -8.4 %, -9.0 %, -10.4 % respectively. Conclusion : The attenuation was caused by presence of side rails and thickness of vac-lok. Pay attention to these attenuation factors, making it a more effective radiation therapy.

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The complete chloroplast genome of Scrophularia kakudensis and a comparative analysis of S. kakudensis and S. cephalantha

  • Ogyeong SON;KyoungSu CHOI
    • Korean Journal of Plant Taxonomy
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    • v.53 no.3
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    • pp.237-241
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    • 2023
  • The genus Scrophularia L. (Scrophulariaceae) comprises 200-270 species worldwide and is a taxonomically challenging lineage, displaying morphological diversity and hybridization. S. kakudensis is morphologically similar to the closely related taxa S. kakudensis var. microphylla, S. pilosa, and S. cephalantha. Therefore, the purpose of this study was to sequence the chloroplast (cp) genome of S. kakudensis using next-generation sequencing and compare it to those of related taxa. The complete cp genome sequence of Scrophularia kakudensis was found to be 152,355 bp long, consisting of a pair of inverted repeats of 25,485 bp that separate a large single-copy (LSC) of 83,479 bp from small single-copy regions of 17,909 bp. The cp genome contained 78 protein-coding genes, 30 tRNAs, and four rRNAs. A phylogenetic analysis based on 78 protein-coding genes from six Scrophularia species showed S. kakudensis and S. cephalantha formed with 100% bootstrap values. We compared the complete cp genomes of S. kakudensis and S. cephalantha and identified seven sequence divergence regions: matK/rps16, rps16/trnQ, trnS/trnG, rpoB/trnC, trnS/trnG, rpl32/trnL, and ndhD/psaC. These regions may be useful for determining the phylogenetic relationships among S. kakudensis-related species.