• 제목/요약/키워드: Sequence Analyses

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Phylogenetic Relationships of Korean Campanulaceae Based on PCR-RFLP and ITS Sequences (PCR-RFLP와 ITS 염기서열 분석을 이용한 한국산 초롱꽃과(Campanulaceae)의 계통유연관계)

  • Kim, Kyung-Ah;Yoo, Ki-Oug
    • Korean Journal of Plant Taxonomy
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    • v.41 no.2
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    • pp.119-129
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    • 2011
  • Phylogenetic studies were conducted to evaluate the taxonomic relationships among 27 taxa, including 2 outgroups of Korean Campanulaceae, using PCR-RFLP analysis and ITS sequences. In the PCR-RFLP analysis, 15 restriction endonucleases produced 244 restriction sites and size variations from the chloroplast DNA, and 59 restriction sites (24%) showed polymorphism. The length of the ITS regions ranged from 588 bp to 797 bp. The sequence divergence including the outgroups is 0-39.36%. Phylogenetic analyses based on PCR-RFLP and ITS data suggest that Campanulaceae is monophyletic; Codonopsis and Platycodon forms an independent clade; the Peracarpa and Asyneuma clade is a sister to the Adenophora-Hanabusaya clade; Campanula is monophyletic; and Wahlenbergia basally branches within the ITS tree, whereas they are placed between Campanula and the Codonopsis-Platycodon clade in the PCR-RFLP tree; Hanabusaya is placed within the Adenophora clade; and Adenophora is paraphyletic and shows discordance to the infrageneric classifications based on morphological data. The present results show two data sets, largely congruent at the generic level, but their phylogenetic positions, in particular the Wahlenbergia and Hanabusaya and the infrageneric classifications in Adenophora, show some incongruence.

The Identity of Eggplant Powdery Mildews Collected in Korea (한국에서 채집된 가지 흰가루병균의 실체)

  • Cho, Sung-Eun;Choi, In-Young;Shin, Hyeon-Dong
    • The Korean Journal of Mycology
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    • v.45 no.2
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    • pp.91-101
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    • 2017
  • Three species of powdery mildew (Erysiphales) on eggplant (Solanum melongena L.) have been listed in Korea, namely Erysiphe cichoracearum (now genus Golovinomyces), Leveillula taurica, and Sphaerotheca fusca (now genus Podosphaera; syn. Podosphaera xanthii). Since E. cichoracearum was recorded on eggplant for the first time in Korea in 1969, it has been regarded as a major powdery mildew agent on that plant. In 1998, the causal agent of powdery mildew on eggplant was recorded as L. taurica, then as S. fusca in 2002. During our extensive field surveys in Korea, we collected 22 samples of eggplant powdery mildews. Our microscopic observations and molecular sequence analyses showed that all of our samples belonged to the genus Podosphaera, in the absence of either E. cichoracearum or L. taurica, suggesting that P. xanthii is the dominant agent of powdery mildew disease on eggplants in Korea. As there have been no additional findings on L. taurica after the first report on the species, it seems to be a minor species that is rarely found in greenhouses. The presence of E. cichoracearum (syn. Golovinomyces cichoracearum s. lat.) on eggplants is questionable, as the morphological characteristics of E. cichoracearum in the original description of the Korean collection deviate from the morphological variations of this species. In addition, no herbarium material of E. cichoracearum remains. Consequently, it seems that P. xanthii is the main species of powdery mildew on eggplants, whereas L. taurica occurs rarely on eggplants, in Korea. This review provides the historical and recent taxonomy of eggplant powdery mildews in detail.

Characterization of the genomes of Aujeszky's disease virus isolated in Korea (국내분리 오제스키병 바이러스의 게놈 유전자 특성 분석)

  • Hyun, Bang-Hun;Kim, In-Joong;Pyo, Hyun-Mi;Cha, Sang-Ho;Park, Ji-Yeun;Song, Jae-Young;Cho, In-Soo;Yang, Chang-Bum;An, Soo-Hwan;Lee, Joong-Bok
    • Korean Journal of Veterinary Research
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    • v.49 no.1
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    • pp.45-57
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    • 2009
  • The molecular genetic characterization of Aujeszky's disease virus (ADV) Yangsan strain (ADVYS), a Korean isolate, was investigated by analyzing the electrophoresis patterns and the physical maps of the viral DNA digested with various endonucleases. To establish DNA library for ADV-YS, twelve major BamHI restricted segments were cloned. Each location of the segments in the ADV genome was determined by sequence comparison with the sequences reported in Genbank and those sequences of the both termini of the segments. Physical maps were constructed based on the electrophoresis patterns of the digested viral DNA by restriction endonuclease and the results of Southern blot analyses with various DIG labeled probes originated from those of enzyme restricted segments of virulent (Shope) and avirulent (Bartha) strain. Comparing ADV-YS with a standard strain of Kaplan in the maps of restriction enzymes, following major respects were identified: (i) disappearance of BamHI restriction site between the first and second BamHI segments, (ii) creation of the BamHI restriction site in the fifth segment, and (iii) generation of the BglII site in the unique short (US) region. The genome of ADV-YS also contains a type 2 herpesvirus DNA molecule (in which the US region only inverts itself relative to the unique longregion) like all other ADV strains except Norden strain(type3), analyzed up to date. The size of the ADV genome estimated from the sizes of the restriction enzyme fragments, was approximately 145.3 kb (BamHI) or 145.4 kb (BglII). BamHI enzyme cleavage patterns were compared among the five Korean ADV isolates: Yangsan, Yongin, Dangjin, Jincheon and Iksan strains. Difference either in the number or in the size of the DNA fragments, suspected regions of termini of IR and TR, could be detected among all five strains.

A LuxR-type Transcriptional Regulator, PsyR, Coordinates Regulation of Pathogenesis-related Genes in Pseudomonas syringae pv. tabaci (Pseudomonas syringae pv. tabaci 에서 LuxR-type 전사조절자인 PsyR에 의한 병원성 유전자들의 조절)

  • Choi, Yeon Hee;Lee, Jun Seung;Yun, Sora;Baik, Hyung Suk
    • Journal of Life Science
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    • v.25 no.2
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    • pp.136-150
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    • 2015
  • Pseudomonas syringae pathovar tabaci is a plant pathogenic bacterium that causes wildfire disease in tobacco plants. In P. syringae pv. tabaci, PsyI, a LuxI-type protein, acts as an AHL synthase, while primary and secondary sequence analysis of PsyR has revealed that it is a homolog of the LuxR-type transcriptional regulator that responds to AHL molecules. In this study, using phenotypic and genetic analyses in P. syringae pv. tabaci, we show the effect of PsyR protein as a quorum-sensing (QS) transcriptional regulator. Regulatory effects of PsyR on swarming motility and production of siderophores, tabtoxin, and N-acyl homoserine lactones were examined via phenotypic assays, and confirmed by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Further qRT-PCR showed that PsyR regulates expression of these virulence genes in response to environmental signals. However, an upstream region of the gene was not bound with purified MBP-PsyR protein; rather, PsyR was only able to shift the upstream region of psyI. These results suggested that PsyR may be indirectly controlled via intermediate-regulatory systems and that auto-regulation by PsyR does not occur.

In vitro Antimicrobial Activity of a New Isolate Streptomyces sp. BCNU 1030 (신규 분리균주 Streptomyces sp. BCNU 1030의 in vitro 항균활성)

  • Bang, Ji-Hun;Choi, Hye-Jung;Ahn, Cheol-Soo;Kim, Dong-Wan;Jeong, Yong-Kee;Joo, Woo-Hong
    • Journal of Life Science
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    • v.21 no.4
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    • pp.589-595
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    • 2011
  • This work focused on screening and characterizing antibiotic-producing actinomycetes to develop new antibiotics that can overcome the growing resistance of disease-causing microbes. One-hundred actinomycetes strains were isolated from soil samples from Chungcheongbuk-do, Korea using various kinds of actinomycetes isolation media, including a starch casein agar medium and potato dextrose agar (PDA). Among them, strain BCNU 1030 was determined to show strong antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA). Biochemical, physiological, and 16S rRNA sequence analyses indicated that strain BCNU 1030 belonged to the genus Streptomyces. Strain BCNU 1030 exhibited antibiotic activity against a wide range of bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentration (MIC) of BCNU 1030 dichloromethane extract was determined to be $0.78\;{\mu}g/ml$ for MRSA CCARM 3090. Therefore, Streptomyces sp. BCNU 1030 has potential for anti-MRSA drug development.

Magnetic Parameters as Indicators of Late-Quaternary Environments on Fort Riley Kansas (암석 자기 변수들을 이용한 제4기 고환경 복원-Fort Riley 캔사스)

  • Park, kyeong
    • The Korean Journal of Quaternary Research
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    • v.11 no.1
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    • pp.57-68
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    • 1997
  • Climatic change of the late-Quaternary period has been record-ed in the loess deposits of the central Great plains and the record of such change is extractable using a number of approaches and parameters. The stratigraphy of loess deposits which have been investigated on Fort Riley exhibits the same sequence of loess units and intercalated buried soils as is found elsewhere in the re-gion but adds detail unique to the reservation Upland late-Qua-ternary composite stratigraphy preserved on the reservation con-sists of the basal Sangamon soil of the Last interglacial(c. 120-110ka), Gilman Canyon Formation(c. >40 -20ka), Peoria loess(c. 20 -10ka) Brady soil(c. 11 -10ka) Bignell loess(c. 9-\ulcornerka). and mod-ern surface soil. Application of magnetic analyses has provided proxy data sets that represent a time series of climatically regulated pedogenesis/weathering and botanical composition. magetic data have yielded an impression of the variation in climate from Sangamon time to the late Holocene through a reconstruction of the history of pedogenesis/weathering. Sangamon soil formation dominated the reservation durin the Last interglacial as indicated by magnetic parameters. During Gil-man Canyon time loess influx was usually sufficiently slow as to permit pedogenesis which appears to have been at a maximum twice during that time. Warm season grasses were important dur-ing soil formation but diminished in importance during the peri-ods of more rapid loess fall which were cooler and perhaps wet-ter. Peoria loess fall a function of the deterioration of climate during the last Glacial Maximum thinly blanketed the reservation with thickest accumulations occurring to the north-west(Bala Cemetery site)proximal to the source region. Long-term surface stability did not apparently occur within Peoria time but short-term stability may be indicaed by the presence of thin weathering zones(incipient soils) in the Peoria loess. Re-gional landscape stability prevailed during the environmental shift at the Pleistocene/Holocene transition resulting in forma-tion of the well expressed Brady soil. One or more weak soils developed in the Bignell loess as it ac-cumulated. A notable feature of the Bignell loess is the appear-ance of the Altithermal dry period: the loess experienced little weathering and was dominated by warm season grasses until the latter of the Holocene.

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Salinity Stress Resistance Offered by Endophytic Fungal Interaction Between Penicillium minioluteum LHL09 and Glycine max. L

  • Khan, Abdul Latif;Hamayun, Muhammad;Ahmad, Nadeem;Hussain, Javid;Kang, Sang-Mo;Kim, Yoon-Ha;Adnan, Muhammad;Tang, Dong-Sheng;Waqas, Muhammad;Radhakrishnan, Ramalingam;Hwang, Young-Hyun;Lee, In-Jung
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.893-902
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    • 2011
  • Endophytic fungi are little known for their role in gibberellins (GAs) synthesis and abiotic stress resistance in crop plants. We isolated 10 endophytes from the roots of field-grown soybean and screened their culture filtrates (CF) on the GAs biosynthesis mutant rice line - Waito-C. CF bioassay showed that endophyte GMH-1B significantly promoted the growth of Waito-C compared with controls. GMH-1B was identified as Penicillium minioluteum LHL09 on the basis of ITS regions rDNA sequence homology and phylogenetic analyses. GC/MS-SIM analysis of CF of P. minioluteum revealed the presence of bioactive $GA_4$ and $GA_7$. In endophyte-soybean plant interaction, P. minioluteum association significantly promoted growth characteristics (shoot length, shoot fresh and dry biomasses, chlorophyll content, and leaf area) and nitrogen assimilation, with and without sodium chloride (NaCl)-induced salinity (70 and 140 mM) stress, as compared with control. Field-emission scanning electron microcopy showed active colonization of endophyte with host plants before and after stress treatments. In response to salinity stress, low endogenous abscisic acid and high salicylic acid accumulation in endophyte-associated plants elucidated the stress mitigation by P. minioluteum. The endophytic fungal symbiosis of P. minioluteum also increased the daidzein and genistein contents in the soybean as compared with control plants, under salt stress. Thus, P. minioluteum ameliorated the adverse effects of abiotic salinity stress and rescued soybean plant growth by influencing biosynthesis of the plant's hormones and flavonoids.

Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library

  • Kim, Dain;Hong, Jisu;Choi, Yoonjoo;Han, Jemin;Kim, Sangkyu;Jo, Gyunghee;Yoon, Jun-Yeol;Chae, Heesu;Yoon, Hyeseon;Lee, Chankyu;Hong, Hyo Jeong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1760-1768
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    • 2020
  • Vibrio cholerae, cause of the life-threatening diarrheal disease cholera, can be divided into different serogroups based on the structure of its lipopolysaccharide (LPS), which consists of lipid-A, core-polysaccharide and O-antigen polysaccharide (O-PS). The O1 serogroup, the predominant cause of cholera, includes two major serotypes, Inaba and Ogawa. These serotypes are differentiated by the presence of a single 2-O-methyl group in the upstream terminal perosamine of the Ogawa O-PS, which is absent in the Inaba O-PS. To ensure the consistent quality and efficacy of the current cholera vaccines, accurate measurement and characterization of each of these two serotypes is highly important. In this study, we efficiently screened a phage-displayed human synthetic Fab library by bio-panning against Ogawa LPS and finally selected three unique mAbs (D9, E11, and F7) that specifically react with Ogawa LPS. The mAbs bound to Vibrio cholerae vaccine in a dose-dependent fashion. Sequence and structure analyses of antibody paratopes suggest that IgG D9 might have the same fine specificity as that of the murine mAbs, which were shown to bind to the upstream terminal perosamine of Ogawa O-PS, whereas IgGs F7 and E11 showed some different characteristics in the paratopes. To our knowledge, this study is the first to demonstrate the generation of Ogawa-specific mAbs using phage display technology. The mAbs will be useful for identification and quantification of Ogawa LPS in multivalent V. cholerae vaccines.

Comparison of 3D Volumetric Subtraction Technique and 2D Dynamic Contrast Enhancement Technique in the Evaluation of Contrast Enhancement for Diagnosing Cushing's Disease

  • Park, Yae Won;Kim, Ha Yan;Lee, Ho-Joon;Kim, Se Hoon;Kim, Sun-Ho;Ahn, Sung Soo;Kim, Jinna;Lee, Seung-Koo
    • Investigative Magnetic Resonance Imaging
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    • v.22 no.2
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    • pp.102-109
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    • 2018
  • Purpose: The purpose of this study is to compare the performance of the T1 3D subtraction technique and the conventional 2D dynamic contrast enhancement (DCE) technique in diagnosing Cushing's disease. Materials and Methods: Twelve patients with clinically and biochemically proven Cushing's disease were included in the study. In addition, 23 patients with a Rathke's cleft cyst (RCC) diagnosed on an MRI with normal pituitary hormone levels were included as a control, to prevent non-blinded positive results. Postcontrast T1 3D fast spin echo (FSE) images were acquired after DCE images in 3T MRI and image subtraction of pre- and postcontrast T1 3D FSE images were performed. Inter-observer agreement, interpretation time, multiobserver receiver operating characteristic (ROC), and net benefit analyses were performed to compare 2D DCE and T1 3D subtraction techniques. Results: Inter-observer agreement for a visual scale of contrast enhancement was poor in DCE (${\kappa}=0.57$) and good in T1 3D subtraction images (${\kappa}=0.75$). The time taken for determining contrast-enhancement in pituitary lesions was significantly shorter in the T1 3D subtraction images compared to the DCE sequence (P < 0.05). ROC values demonstrated increased reader confidence range with T1 3D subtraction images (95% confidence interval [CI]: 0.94-1.00) compared with DCE (95% CI: 0.70-0.92) (P < 0.01). The net benefit effect of T1 3D subtraction images over DCE was 0.34 (95% CI: 0.12-0.56). For Cushing's disease, both reviewers misclassified one case as a nonenhancing lesion on the DCE images, while no cases were misclassified on T1 3D subtraction images. Conclusion: The T1 3D subtraction technique shows superior performance for determining the presence of enhancement on pituitary lesions compared with conventional DCE techniques, which may aid in diagnosing Cushing's disease.

Repression of HspA2 mRNA Expression in Human Testes with Abnormal Spermatogenesis (비정상적 정자형성 환자의 정소에서 Heat Shock Protein A2 (hspA2) mRNA 발현의 감소)

  • Son, W.Y.;Hwang, S.H.;Han, C.T.;Lee, J.H.;Kim, S.J.;Kim, Y.C.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.103-109
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    • 1999
  • Objective: Heat shock protein 70-2 (Hsp70-2) gene knockout mice are found to have premeiotic arrest at the primary spermatocyte stage with a complete absence of spermatids and spermatozoa. This observation led to the hypothesis that hspA2 may be disrupted in human testes with abnormal spermatogenesis. To test this hypothesis, we studied the mRNA expression of hspA2 in infertile men with azoospermia. Design: The mRNA expression were analyzed by competitive RT-PCR among testes with normal spermatogenesis, pachytene spermatocyte arrest, and sertoli-cell only syndrome. Materials and methods: Testicular biopsy was performed in men with azoospermia (n=15). Specimens were subdivided into three groups: (group 1) normal spermatogenesis (n=5), (group 2) spermatocyte arrest (n=5), (group 3) Sertoli-cell only syndrome (n=5). Total RNA was extracted by Trizol reagent. Total extracted RNA was reverse transcribed into cDNA and amplified by PCR using specific primers for hspA2 target cDNAs. A competitive cDNA fragment was constructed by deleting a defined fragment from the target cDNA sequence, and then coamplified with the target cDNA for competitive PCR. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal control. Results: On Competitive RT-PCR analyses for hspA2 mRNA, significant amount of hspA2 expression was observed in group 1, whereas a constitutively low level of hspA2 was expressed in groups 2 and 3. Conclusion(s): The study demonstrates that the hspA2 gene expression is down-regulated in human testes with abnormal spermatogenesis, which in turn suggests that hspA2 gene may play a specific role during meiosis in human testes.

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