• Title/Summary/Keyword: DNA 염기

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Protease Activity of Lactic Acid Bacteria Isolated from Korean Traditional Fermented Food (전통 발효식품으로부터 Protease 활성을 보유한 유산균의 분리 및 동정)

  • Kook, Moo Chang;Cho, Seok Cheol;Park, Hoon;Kim, Seung Seop;Pyun, Yu Ryang;Choi, Woon Yong;Lee, Hyeon Yong
    • Food Engineering Progress
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    • v.15 no.2
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    • pp.182-187
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    • 2011
  • A proteolytic lactic acid bacterium was isolated from Korean traditional fermented foods. The isolate BV-26, which had a protease activity (24 U/mg-crude protein), was identified as Lactobacillus plantarum by the API 50CHL kit and 16S rDNA analysis (99.9% of homology), and named as L. plantarum BV-26. Cell growth and protease activity of L. plantarum BV-26 was determined in MRS broth using 5L jar fermentor at $30^{\circ}C$. The maximum growth of L. plantarum BV-26 was reached at 18 hr in MRS broth, while protease activity of BV-26 was detectable at 12 hr and the highest activity was obtained after 16 hr cultivation. Therefore, we expect that the proteolytic lactic acid bacteria, L. plantarum BV-26, may be used as a starter for the fermentation of animal feed. Especially, the fermentation of soybean meal with the strain can be applied for improving feed utilization.

Breeding of F1 Hybrid for Oriental Mustard(Brassica juncea L. Czern) Using the Cytoplasmic Male Sterile Line (웅성불임 인자를 이용한 갓(Brassica juncea L. Czern)의 F1 육종)

  • Park, Yong Ju;Min, Byung Whan
    • Journal of agriculture & life science
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    • v.52 no.6
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    • pp.27-36
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    • 2018
  • Recently, the demand for new cultivar of oriental mustard(Brassica juncea) is increased as the consumption of oriental mustard has increased dramatically in the market due to Kimchi is attracting the world's attention. However, absence of seed supply system can cause many problems including inbreeding depression due to self seed production, deterioration of the seed purity and heterogeneity of commercial seed. To establish the $F_1$ variety breeding system of oriental mustard, pure line and inbred lines were screened from inbreeding genetic resources. Male-sterile line was also selected from breeding combinations for the quality improvement of mustard. The combining ability from(Indojasai ${\times}$ Goheungdamyang) combinations and isolation line of(MS910 ${\times}$ Japan red mustard 8 ${\times}$ Ganghwa mustard 9) was highest, thus these lines were selected as parental lines. PCR(Polymerase Chain Reaction) and gene sequence analysis revealed that the genes related to CMS(orf263, orf220, and orf288) were distributed in mitochondria. The isolated lines from this study also showed good performance in yield test and farmhouse prove test.

Molecular Analysis of Carbapenem-Resistant Pseudomonas aeruginosa Isolated from Patients Hospitalized in Daejeon between 2008 and 2014 Years (대전지역의 입원환자에서 분리된 Carbapenem 내성 Pseudomonas aeruginosa의 분자역학조사(2008년에서 2014년까지))

  • Cho, Hye Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.406-413
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    • 2018
  • The emergence of carbapenem resistance among Pseudomonas aeruginosa has become an increasing problem worldwide. In particular, $metallo-{\beta}-lactamases$ (MBLs) are responsible for the high-level resistance to carbapenem. Sequence type 235 (ST235) has been found internationally in a multidrug-resistant clone and is involved in the dissemination of genes encoding IMP-6 and VIM-2. This study examined the prevalence of MBLs and the epidemiological relationship in carbapenem-resistant P. aeruginosa (CRPA) isolates obtained from a tertiary hospital in Daejeon, Korea, between March 2008 and June 2014. The antimicrobial susceptibilities were determined using the disk-diffusion method and PCR and DNA sequencing were used to identify the MBL genes. In addition, an epidemiological relationship was investigated by multilocus sequence typing (MLST). Among the 110 CRPA isolates, 32 isolates (29.1%) were MBL-producers; the major type was IMP-6 (29 isolates, 90.6%). VIM-2 was identified in 3 isolates (9.4%) of ST357. IMP-6-producing isolates were multidrug-resistant (MDR) and belonged to ST235. ST235 (55 isolates, 50.0%) was the clone most frequently detected and has gradually emerged during a seven-year period. To prevent the spread of MDR ST235 P. aeruginosa isolates, the current widespread use of carbapenems needs to be curtailed, and novel continuous monitoring strategies should be developed as soon as possible.

Isolation and Identification of Ampicillin-resistant Bacteria in Changwon (창원근교에서의 ampicillin 내성세균의 분리 및 동정)

  • Bae, Young-Min
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1529-1535
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    • 2018
  • The number of antibiotic-resistant bacteria is increasing rapidly while the discovery rate of new antibiotics is in decline. A systematic study is therefore necessary to investigate which bacteria are resistant to medically important antibiotics and how high that resistance is. To that end, this study aimed to analyze which bacteria demonstrated resistance to ampicillin, one of the currently most-widely used medical antibiotics. Water samples were collected from the Changwon-Cheon that runs through Changwon City and from the pond in front of the dormitory building at Changwon University. Hundreds of ampicillin-resistant colonies were obtained and 22 morphologically distinct examples were chosen for further study. These bacteria were identified by amplifying their 16S rRNA genes and comparing those sequences with data in GenBank. The bacteria was identified as belonging to 10 families, 12 genera, and 17 species, and all were able to grow in the presence of $50{\mu}g/ml$ ampicillin while seven showed growth at ampicillin concentrations as high as 1.5 mg/ml.

Molecular Detection of Virulence Factors in Carbapenem-Resistant Pseudomonas aeruginosa Isolated from a Tertiary Hospital in Daejeon (대전지역의 3차 병원에서 분리된 Carbapenem 내성 Pseudomonas aeruginosa의 병독성 인자 검출)

  • Cho, Hye Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.301-308
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    • 2019
  • The emergence and spread of multidrug resistant (MDR) Pseudomonas aeruginosa is a critical problem worldwide. The pathogenesis of P. aeruginosa is due partly to the production of several cell-associated and extracellular virulence factors. This study examined the distribution of virulence factors and antimicrobial resistance patterns of carbapenem-resistant P. aeruginosa (CRPA) isolated from a tertiary hospital in Daejeon, Korea. Antimicrobial susceptibility testing was performed using the disk diffusion method, and PCR and DNA sequencing were performed to determine for the presence of virulence genes. In addition, the sequence type (ST) of MDR P. aeruginosa was investigated by multilocus sequence typing (MLST). Among 32 CRPA isolates, 14 (43.8%) were MDR and the major ST was ST235 (10 isolates, 71.4%). All isolates were positive for the presence of virulence genes and the most prevalent virulence genes were toxA, plcN, and phzM (100%). All isolates carried at least eight or more different virulence genes and nine (28.1%) isolates had 15 virulence genes. The presence of the exoU gene was detected in 71.4% of the MDR P. aeruginosa isolates. These results indicate that the presence of the exoU gene can be a predictive marker for the persistence of MDR P. aeruginosa isolates.

Isolation of the Protease-producing Yeast Pichia anomala CO-1 and Characterization of Its Extracellular Neutral Protease (세포 외 중성 단백질분해효소를 생산하는 Pichia anomala CO-1의 분리 동정 및 효소 특성)

  • Kim, Ji Yeon
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1126-1135
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    • 2019
  • From a sample of bamboo byproduct, the protease-producing yeast strain CO-1 was newly isolated. Strain CO-1 is spherical to ovoid in shape and measures $3.1-4.0{\times}3.8-4.4{\mu}m$. For the growth of strain CO-1, the optimal temperature and initial pH were $30^{\circ}C$ and 4.0, respectively. The strain was able to grow in 0.0-15.0%(w/v) NaCl and 0.0-9.0%(v/v) ethanol. Based on a phylogenetic analysis of its 18S rDNA sequences, strain CO-1 was identified as Pichia anomala. The extracellular protease produced by P. anomala CO-1 was partially purified by ammonium sulfate precipitation, which resulted in a 14.6-fold purification and a yield of 7.2%. The molecular mass of the protease was recorded as approximately 30 kDa via zymogram. The protease activity reached its maximum when 1.0%(w/v) CMC was used as the carbon source, 1.0%(w/v) yeast extract was used as the nitrogen source, and 0.3%(w/v) $MnSO_4$ was used as the mineral source. The protease revealed the highest activity at pH 7.0 and $30^{\circ}C$. This enzyme maintained more than 75% of its stability at a pH range of 4.0-10.0. After heating at $65^{\circ}C$ for 1 hr, the neutral protease registered at 60% of its original activity. The protease production coincided with growth and attained a maximal level during the post-exponential phase.

A Study of the Diversity and Profile for Extracellular Enzyme Production of Aerobically Cultured Bacteria in the Gut of Muraenesox cinereus (갯장어(Muraenesox cinereus) 장으로부터 호기적 조건에서 분리된 미생물의 다양성 및 세포외 효소 생산능 분석에 관한 연구)

  • Lee, Yong-Jik;Oh, Do-Kyoung;Kim, Hye Won;Nam, Gae-Won;Sohn, Jae Hak;Lee, Han-Seung;Shin, Kee-Sun;Lee, Sang-Jae
    • Journal of Life Science
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    • v.29 no.2
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    • pp.248-255
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    • 2019
  • This research confirmed the diversity and characterization of gut microorganisms isolated from the intestinal organs of Muraenesox cinereus, collected on the Samcheonpo Coast and Seocheon Coast in South Korea. To isolate strains, Marine agar medium was basically used and cultivated at $37^{\circ}C$ and pH7 for several days aerobically. After single colony isolation, totally 49 pure single-colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rRNA gene DNA sequencing, indicating that isolated strains were divided into 3 phyla, 13 families, 15 genera, 34 species and 49 strains. Proteobacteria phylum, the main phyletic group, comprised 83.7% with 8 families, 8 genera and 26 species of Aeromonadaceae, Pseudoalteromonadaceae, Shewanellaceae, Enterobacteriaceae, Morganellaceae, Moraxellaceae, Pseudomonadaceae, and Vibrionaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 39 strains possessed at least one enzyme activity. Especially the Aeromonas sp. strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed for securing domestic genetic resources and the expansion of scientific knowledge of the gut microbial community in Muraenesox cinereus of South Korea.

Distribution of Subgroups in Pseudomonas syringae pv. actinidiae Biovar 3 Strains Isolated from Korea (국내에서 분리된 Pseudomonas syringae pv. actinidiae biovar 3 균주들의 subgroup 분포)

  • Lee, Young Sun;Kim, Gyoung Hee;Jung, Jae Sung
    • Journal of Life Science
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    • v.31 no.1
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    • pp.52-58
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    • 2021
  • Pseudomonas syringae pv. actinidiae, which causes bacterial canker in kiwifruit, is divided into five biovars (1, 2, 3, 5, 6) on the basis of genetic characteristics and toxin productivity. Among them, biovar 3 is responsible for the current global outbreak, and has been isolated in Korea since 2011. Biovar 3 strains isolated from Korea are subdivided into six genetically different lineages (subgroup I, IV, V, VI, VII, and VIII) based on random amplified polymorphic DNA (RAPD) analysis. In this work, the subgroup-specific sequence characterized amplified region (SCAR) primers were developed from sequenced differential RAPD bands. Distribution of the subgroups of the biovar 3 strains collected in Korea from 2011-2017 were examined using these subgroup-specific primer sets. Among the 54 strains tested, 35 strains (64.8%) belonged to subgroup V, 9 strains (16.7%) belonged to subgroup IV, 4 strains (7.4%) belonged to subgroup VI, 3 strains (5.6%) belonged to subgroup VII, 2 strains (3.7%) belonged to subgroup VIII, and 1 (1.9%) strain belonged to subgroup I. Strains belonging to subgroups IV, V, and VI were shown to be related to strains isolated from China, New Zealand, and Chile, respectively. The study revealed that the biovar 3 strains in Korea are genetically diverse and are estimated to have been introduced through pollen sourced from foreign countries.

Characterization of Three Korean Isolates of Malva Vein Clearing Virus from Curled Mallow (Malva verticillata) (아욱에서 분리한 Malva Vein Clearing Virus 분리주의 특성)

  • Kwak, Hae-Ryun;Kim, Ji-Gwang;Kim, Jeong-Eun;Choi, Hyeon-Yong;Choi, Hong-Soo;Kim, Mikyeong
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.283-288
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    • 2020
  • In September 2017, vein clearing and yellowing symptoms resembling those caused by viruses were observed on leaves of Malva verticillata in Chungnam, Korea. Nucleic acids were extracted from leaves of five symptomatic plants and tested by reverse transcription polymerase chain reaction using four virus specific primer pairs including malva vein clearing virus (MVCV). Amplicons of the expected size (600 bp) were obtained from total RNA of all samples using the MVCV-specific primers. To confirm the presence of MVCV in symptomatic plants, the DNA fragments from three samples were purified, and directly sequenced. BLAST analysis revealed that it shared the highest nucleotide identity (99%) with a MVCV isolate from tomato (Mexico). The virus isolates obtained from the third re-inoculated Chenopodium was designated as Cm1-5. Tissue from Cm1, Cm3, and Cm5 isolates was mechanically sap inoculated into 23 indicator plants. Cm3 isolate induced chlorotic local and mosaic symptoms in Althaea rosea. Phylogenetic analysis based on coat protein gene of 19 MVCV isolates from 6 different countries and plant species, did not correlated with either the geographical origin of the isolates, or pathogenicity. To our knowledge, this study first reports the natural occurrence of MVCV on M. verticillata in Korea and characterization of three Korean isolates of MVCV.

Development of ITS sequence based SCAR marker and multiplex-SCAR assay for the rapid authentication of Tetrapanacis Medulla and Akebiae Caulis (통초(通草), 목통(木通) 신속 감별용 ITS 염기서열 기반 SCAR 마커 및 Multiplex-SCAR 분석법 개발)

  • Noh, Pureum;Kim, Wook Jin;Park, Inkyu;Yang, Sungyu;Choi, Goya;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.36 no.1
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    • pp.9-17
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    • 2021
  • Objectives : Tetrapanacis Medulla and Akebiae Caulis are one of the most frequently adulterated herbal medicines because of their confusability of terms in the ancient writings and the similarity of morphological features of dried herbal products. The major adulterant is Aristolochia manshuriensis (Guanmutong) which has a serious safety concern with its toxicity. To ensure the safety and quality of the two herbal medicines, it is necessary to discriminate the toxic adulterant from authentic species. The aim of this study is to develop SCAR markers and to establish the multiplex-SCAR assay for discrimination of four plant species related to Tetrapanacis Medulla and Akebiae Caulis. Methods : ITS regions of fifteen samples of four species (Tetrapanax papyrifer, Fatsia japonica, Aristolochia manshuriensis, and Akebia quinata) collected from different sites were amplified and sequenced. Fifteen obtained ITS sequences were aligned and analysed for the detection of species-specific sequence variations. The SCAR markers were designed based on the sequence alignments and then, multiplex-SCAR assay enhancing rapidity was optimized. Results : ITS sequences clearly distinguished the four species at the species level. The developed SCAR markers and multiplex-SCAR assay were successfully discriminated four species and detected the adulteration of commercial product samples by comparison of the amplified DNA fragment sizes. Conclusions : These SCAR markers and multiplex-SCAR assay are a rapid, simple, and reliable method to identify the authentic Tetrapanacis Medulla and Akebiae Caulis from adulterants. These genetic tools will be useful to ensure the safety and to standardize the quality of the two herbal medicines.