• 제목/요약/키워드: molecular profile

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Xanthomonas axonopodis pv. eucalyptorum pv. nov. Causing Bacterial Leaf Blight on Eucalypt in Brazil

  • Ferraz, Helvio Gledson Maciel;Badel, Jorge Luis;da Silva Guimaraes, Lucio Mauro;Reis, Bruna Paolinelli;Totola, Marcos Rogerio;Goncalves, Rivadalve Coelho;Alfenas, Acelino Couto
    • The Plant Pathology Journal
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    • v.34 no.4
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    • pp.269-285
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    • 2018
  • Bacterial leaf blight is a major disease of eucalypt, especially under nursery conditions. Different bacterial species have been associated with the disease in several countries, and despite its importance worldwide, it is not clear to date whether similar disease symptoms are caused by the same or by different etiological agents. In this study, 43 bacterial strains were isolated from blighted eucalypt leaves collected in different geographic areas of Brazil and inoculated onto a susceptible eucalypt clone. Polyphasic taxonomy, including morphological, physiological, biochemical, molecular, and pathogenicity tests showed that only certain strains of Xanthomonas axonopodis caused symptoms of the disease. Strains varied in their aggressiveness, but no correlation with geographic origin was observed. MLSA-based phylogenetic analysis using concatenated dnaK, fyuA, gyrB and rpoD gene sequences allocated the strains in a well-defined clade, corresponding to Rademarker's group RG 9.6. Inoculation of nineteen plant species belonging to seven botanical families with representative strain LPF 602 showed it to be pathogenic only on Eucalyptus spp, and Corymbia spp. Based on distinct biochemical and pathogenic characteristics that differentiate the eucalypt strains from other pathovars of the X. axonopodis species, here we propose their allocation into the new pathovar X. axonopodis pv. eucalyptorum pv. nov.

Gene expression analysis related to ethylene induced female flowers of cucumber (Cucumis sativus L.) at different photoperiod

  • Ikram, Muhammad Maulana Malikul;Esyanti, Rizkita Rachmi;Dwivany, Fenny Martha
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.229-234
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    • 2017
  • Photoperiod is one of the factors affecting productivity of cucumber plant by inducing ethylene hormone production and so triggering flower sex differentiation into female flower. However, only few studies have been perfomed in order to reveal the effect of photoperiod in molecular level in relation to the flower differentiation. Therefore, in this study, Mercy cultivar of cucumber (andromonoecious) was treated with photoperiod of 8, 12, 16 hours of light, while control received no treatment of additional light. Photoperiod of 8 hours was achieved by blocking the sunlight with shade net and 16 hours by giving longer light exposure using white LEDs. Cucumber's flowers were quantified and the apical and lateral shoots were extracted to evaluate the gene profile related to the photoperiod, ethylene production, and female flower differentiation, which were CsACS2, CsETR1, CsCaN, and CsPIF4 using PCR method. Photoperiod of 8 hours affected the production of female flower with average number of 6.7 flowers in main stem and 8.0 flowers in lateral stem, compared to photoperiod of 12 and 16 hours which produced 3.7 and 2.0 flowers in main stem with 7.0 and 11.3 in lateral stem, respectively. In silico studies in this experiment resulted in proposed model of signal transduction that showed the connection between ethylene production and flower differentiation. PCR analysis confirmed the expression of CsACS2, CsETR1, and CsCaN, that was positively correlated with numbers of female flowers in cucumber, but the expression of CsPIF4 that represent photoperiod haven't been confirmed correlated with the ethylene production and flower differentiation.

Genetic properties of R plasmids in Shigella isolates of swine origin in Korea (돈(豚)에서 분리(分離)한 Shigella균유래(菌由來) R plasmid의 유전적(遺傳的) 특성(特性)에 관한 연구(硏究))

  • Choi, Won-pil;Kwun, Hae-byeng;Jung, Suk-chan
    • Korean Journal of Veterinary Research
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    • v.29 no.1
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    • pp.37-44
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    • 1989
  • This paper dealt with the distribution of Shigella spp. on 5 piggeries in Taegu and Kyungpook during the period from August to October 1987. Isolated Shigella were examined for serogrouping, antimicrobial drug resistance and detection of R plasmid. Genetic properties of R plasmid in Shigella have examined to fertility inhibition (Fi) and gel electrophoresis was performed for the isolation of plasmid DNA. The results obtained were summarised as followings; 1. Of total 2,978 samples from 5 piggeries, 82 strains (2.8%) of Shigella spp. were isolated from 82 samples. The isolated strains were identified as S dysenteriae (60 strains), S flexneri (20 strains) and S sonnei (2 strains). 2. Of the 82 strains examined 67 (95.1%) were resistant to one or more antibiotics, such as ampicillin (Am), chloramphenicol (Cm), kanamycin (Km), nalidixic acid (Na), rifampicin (Rf), streptomycin (Sm), sulfademethoxine (Su), and tetracycline (Tc) and higher resistant to Su (90.2%), Sm (63.4%) and Tc (63.4%). 3. Of the 78 resistant Shigella strains 26 (33.3%) harbored conjugative R plasmids and the transfer frequency of Sm (50.0%), Cm(33.3%) resistance was much higher than that of the other drug resistance. 4. The most common resistant patterns were SmSuTc, Su and AmSmSuTc. 5. Out of the 26 Shigella R plasm ids examined for Fi, 14(53.8%) were $Fi^+$ and the remainder were $Fi^-$. 6. The plasmid DNA profiles in Shigella spp. (9 strains) isolated from pigs were confirmed as being 2 to 9 fragments by the gel electrophoresis. Their molecular size ranged 2.17 to 87.62 kilobase (Kb). All strains of Shigella spp. consisted in 15.4 Kb plasmids.

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Gene Profiles in a Smoke-Induced COPD Mouse Lung Model Following Treatment with Mesenchymal Stem Cells

  • Kim, You-Sun;Kokturk, Nurdan;Kim, Ji-Young;Lee, Sei Won;Lim, Jaeyun;Choi, Soo Jin;Oh, Wonil;Oh, Yeon-Mok
    • Molecules and Cells
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    • v.39 no.10
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    • pp.728-733
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    • 2016
  • Mesenchymal stem cells (MSCs) effectively reduce airway inflammation and regenerate the alveolus in cigarette- and elastase-induced chronic obstructive pulmonary disease (COPD) animal models. The effects of stem cells are thought to be paracrine and immune-modulatory because very few stem cells remain in the lung one day after their systemic injection, which has been demonstrated previously. In this report, we analyzed the gene expression profiles to compare mouse lungs with chronic exposure to cigarette smoke with non-exposed lungs. Gene expression profiling was also conducted in a mouse lung tissue with chronic exposure to cigarette smoke following the systemic injection of human cord blood-derived mesenchymal stem cells (hCB-MSCs). Globally, 834 genes were differentially expressed after systemic injection of hCB-MSCs. Seven and 21 genes, respectively, were up-and downregulated on days 1, 4, and 14 after HCB-MSC injection. The Hbb and Hba, genes with oxygen transport and antioxidant functions, were increased on days 1 and 14. A serine protease inhibitor was also increased at a similar time point after injection of hCB-MSCs. Gene Ontology analysis indicated that the levels of genes related to immune responses, metabolic processes, and blood vessel development were altered, indicating host responses after hCB-MSC injection. These gene expression changes suggest that MSCs induce a regeneration mechanism against COPD induced by cigarette smoke. These analyses provide basic data for understanding the regeneration mechanisms promoted by hCB-MSCs in cigarette smoke-induced COPD.

A Simple Method for the Detection and Identification of Korean Brown Cattle(Hanwoo) using DHPLC

  • Kang, Won;Park, Sane-Bum;Nam, Youn-Hyoung;An, Young-Chang;Lee, Sang-Hyun;Park, Jin-Keun;Lee, Jae-Sik;Jang, On-Cheoul;Park, Su-Min;Park, Yong-Hyun;Ko, Kyung-Hee;Moon, Sung-Sil
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2006.05a
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    • pp.179-183
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    • 2006
  • Denaturing high-performance liquid chromatography(DHPLC) is used in a wide variety of genetic applications and is an efficient method for detection of mutations involving one or a few nucleotides. We developed a high-throughput DHPLC method for identifying polymorphisms in the MC1R gene that are characteristic of Hanwoo cattle. We compared 10 tissue samples from Hanwoo cattle, 10 samples from Holstein cattle and 10 samples from Hanwoox Holstein crossbred cattle to determine whether DHPLC analysis can be used to distinguish between these genotypes. Samples obtained from Hanwoo cattle had a unique profile of peaks that could be used as a molecular fingerprint for this breed. We also analyzed two hundred samples in a trial in which we were blinded to the genotype of the samples and correctly identified the breed-of-origin of 594 out of 600 sequence variations(99%).

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Profiling of Disease-Related Metabolites in Grapevine Internode Tissues Infected with Agrobacterium vitis

  • Jung, Sung-Min;Hur, Youn-Young;Preece, John E.;Fiehn, Oliver;Kim, Young-Ho
    • The Plant Pathology Journal
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    • v.32 no.6
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    • pp.489-499
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    • 2016
  • Green shoot cuttings of 10 different grapevine species were inoculated with Agrobacterium vitis to find disease-related metabolites in the grapevine. Crown galls formed 60 days after inoculation varied in gall severity (GS) evaluated by gall incidence (GI) and gall diameter (GD), which were classified into three response types as RR (low GI and small GD), SR (high GI and small GD), and SS (high GI and large GD), corresponding to resistant, moderately resistant, and susceptible responses, respectively. In this, 4, 4, and 2 Vitis species were classified into RR, SR, and SS, respectively. Gas chromatography mass spectrometry (GC-MS) analysis of the grapevine stem metabolites with A. vitis infection showed 134 metabolites in various compound classes critically occurred, which were differentially clustered with the response types by the principal component analysis. Multivariate analysis of the metabolite profile revealed that 11 metabolites increased significantly in relation to the response types, mostly at post-inoculation stages, more prevalently (8 metabolites) at two days after inoculation than other stages, and more related to SS (7 metabolites) than RR (3 metabolites) or SR (one metabolite). This suggests most of the disease-related metabolites may be rarely pre-existing but mostly induced by pathogen infection largely for facilitating gall development except stilbene compound resveratrol, a phytoalexin that may be involved in the resistance response. All of these aspects may be used for the selection of resistant grapevine cultivars and their rootstocks for the control of the crown gall disease of the grapevine.

Cell adaptation of KPEDV-9 and serological survey on porcine epidemic diarrhea virus(PEDV) infection in Korea (돼지 유행성설사바이러스 국내 분리주(KPED-9) 의 세포증식성 및 혈청학적 역학조사)

  • Kweon, Chang-hee;Kwon, Byung-joon;Kang, Yung-bai;An, Soo-hwan
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.321-326
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    • 1994
  • Korean isolate, porcine epidemic diarrhea virus (KPEDV-9) was adapted through serial passages in vero cell. The viral yield reached up to $10^{5-6}$ $TCID_{50}/ml$ at the passage level of 90th. The cell adapted virus was characterized through genetic and morphological examinations. The RNA extracted from virus infected cell revealed the presence of RNA band with molecular size of >20Kb. The electron microscopic examination on purified virus showed the pleomorphic appearance of enveloped particles with 5-10nm surface projections, which fit with the shape of coronavirus. The etiological survey on swine diarrhea by immunofluorescence test(FA) indicated 17.5% positive rate on the PEDV infection. In addition, the incidence were detected both in piglets within two weeks old as well as fattening pigs. Serological survey by ELISA revealed the overall 45% positive result, thus, indicating the PEDV infection are widespread throughout this country.

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A Unique Gene Expression Signature of 5-fluorouracil

  • Kim, Ja-Eun;Yoo, Chang-Hyuk;Park, Dong-Yoon;Lee, Han-Yong;Yoon, Jeong-Ho;Kim, Se-Nyun
    • Molecular & Cellular Toxicology
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    • v.1 no.4
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    • pp.248-255
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    • 2005
  • To understand the response of cancer cells to anticancer drugs at the gene expression level, we examined the gene expression changes in response to five anticancer drugs, 5-fluorouracil, cytarabine, cisplatin, paclitaxel, and cytochalasin D in NCI-H460 human lung cancer cells. Of the five drugs, 5-fluorouracil had the most distinctive gene expression signature. By clustering genes whose expression changed significantly, we identified three clusters with unique gene expression patterns. The first cluster reflected the up-regulation of gene expression by cisplatin, and included genes involved in cell death and DNA repair. The second cluster pointed to a general reduction of gene expression by most of the anticancer drugs tested. A number of genes in this cluster are involved in signal transduction that is important for communication between cells and reception of extracellular signals. The last cluster represented reduced gene expression in response to 5-fluorouracil, the genes involved being implicated in DNA metabolism, the cell cycle, and RNA processing. Since the gene expression signature of 5-fluorouracil was unique, we investigated it in more detail. Significance analysis of microarray data (SAM) identified 808 genes whose expression was significantly altered by 5-fluorouracil. Among the up-regulated genes, those affecting apoptosis were the most noteworthy. The down-regulated genes were mainly associated with transcription-and translation-related processes which are known targets of 5-fluorouracil. These results suggest that the gene expression signature of an anticancer drug is closely related to its physiological action and the response of caner cells.

A Study on Electrophoretic Pattern and Immunosuppressive Activity of Uterine Secretory Protein in Early Pregnant Sows (초기 임신돈에 있어서 자궁분비단백질의 전기영동상과 면역억제력에 관한 연구)

  • 김경순;정영채;김창근;한기영;김형태;윤종택;김종대;최선호
    • Korean Journal of Animal Reproduction
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    • v.13 no.3
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    • pp.179-187
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    • 1989
  • The experiment was carried out to study the profile of uterine specific protein during early pregnancy in sows and to test it's immunosuppressive activity. Uterine protein samples were obtained by flushing the uterine horn on Day 3, 6, 9, 12 and 15 of the estrous cycle and the pregnancy respectively and the protein concentration of each sample was determined. The change of uterine protein was examined by sodium dodecyl sulfate(SDS)-PAGE. The immunosuppressive activity of uterine secretory protein was investigated according to the lymphocyte blastogenesis response to mitogen. The results of this experiment are summarized as follows ; 1. The uterine protein during estrous cycle and early pregnancy was relatively constant up to Day 9, but increased on Day 12. Maxium total protein values were found on Day 15. The concentration of serum proteins were about 82-95 mg during estrous cycle, but decreased to about 70-82 mg during early pregnancy. 2. The proteins components similar electrophoretic patterns(PAGE) that were no differences (band ; a, b, c, d, e, f, g, I) on Days 3, 6 and 9 of the estrous cycle and pregnancy. But there were more 2 bands specifically on Day 12 of the pregnancy and on Day 15 of estrous cycle and showed more 4 bands on Day 15 of early pregnancy. They seemed to be acidoprotein and their average molecular weight were 38,000, 22,300 and 12,600. 3. When uterine protein were added 500$\mu\textrm{g}$/ml, there was no immunosuppresive activity on Day 3 of estrous cycle and lymphocyte blastogenesis was slightly suppressed on Day 3 of pregnancy. The immunosuppressive activity on Day 9 of estrous cycle and pregnancy appeared in 500$\mu\textrm{g}$/ml and 150$\mu\textrm{g}$/ml, respectively the uterine protein on Day 12 and 15 showed immunosuppresive activity, which at the level of 150$\mu\textrm{g}$/ml during non-pregnancy and at the level of 100 to 125$\mu\textrm{g}$/ml during early pregnancy, respectively.

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Effect of irradiation on the Streptococcus mutans (방사선조사가 Streptococcus mutans에 미치는 영향)

  • Ahn, Ki-Dong;Kim, Gyu-Tae;Choi, Yong-Suk;Hwang, Eui-Hwan
    • Imaging Science in Dentistry
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    • v.37 no.1
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    • pp.35-43
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    • 2007
  • Purpose : To observe direct effect of irradiation on cariogenic Streptooccus mutans. Materials and Methods : S. mutans GS5 was exposed to irradiation with a single absorbed dose of 10, 20, 30, and 40Gy. Viability and changes in antibiotic sensitivity, morphology, transcription of virulence factors, and protein profile of bacterium after irradiation were examined by pour plate, disc diffusion method, transmission electron microscopy, RT-PCR, and SDS-PAGE, respectively. Results : After irradiation with 10 and 20Gy, viability of S. mutans was reduced. Further increase in irradiation dose, however, did not affect the viability of the remaining cells of S. mutans. Irradiated 5. mutans was found to have become sensitive to antibiotics. In particular, the bacterium irradiated with 40Gy increased its susceptibility to cefotaxime, penicillin, and tetracycline. Under the transmission electron microscope, number of morphologically abnormal cells was increased as the irradiation dose was increased. S. mutans irradiated with 10 Gy revealed a change in the cell wall and cell membrane. As irradiation dose was increased, a higher number of cells showed thickened cell wall and cell membrane and Iysis, and appearance of ghost cells was noticeable. In RT-PCR, no difference was detected in expression of gtfB and spap between cells with and without irradiation of 40Gy. In SDS-PAGE, proteins with higher molecular masses were gradually diminished as irradiation dose was increased. Conclusion : These results suggest that irradiation affects the cell Integrity of S. mutans, as observed by SDS-PAGE, and as manifested by the change in cell morphology, antibiotic sensitivity, and eventually viability of the bacterium.

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