• Title/Summary/Keyword: PCR 동정

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Sphingobacterium sp. SW-09 Effectively Degrades Phenanthrene, a Polycyclic Aromatic Hydrocarbon, in a Soil Microcosm (Sphingobacterium sp. SW-09에 의한 토양환경에서의 다환 방향족탄화수소인 페난스렌의 분해)

  • Son, Seung-Woo;Chang, Hey-Won;Kim, Sung-Kuk;Chang, Jong-Soo
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1511-1517
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    • 2011
  • We isolated a potent phenanthrene-degrading bacterium from oil-contaminated soils of Suzhou, China, and assessed the potential use of these bacteria for bioremediation of soils contaminated by polycyclic aromatic hydrocarbons (PAHs) in a microcosm. Based on 16S rDNA sequencing, we identified this bacteria as Sphigobacterium sp. SW-09. By PCR amplification, we also identified catechol 2,3-dioxygenase genes (nahH genes) mediating PAH degradation. Staphylococcus sp. KW-07, which has been identified in our previous study, showed potential for use in bioremediation of oil-contaminated soils. In this experiment, we compared the rate of phenanthrene-degradation between Staphylococcus sp. KW-07 and Sphingobacterium sp. SW-09 in a microcosm condition. Newly isolated Sphingobacterium sp. SW-09 showed a higher phenanthrene-degradation rate than that of Staphylococcus sp. KW-07 in soil microcosms. Together, our results suggest that the Sphingobacterim sp. SW-09 strain isolated from the Suzhou area may also be useful in bioremediation of PAH-contaminated soils.

Molecular Phylogenetic Analysis of Botrytis cinerea Occurring in Korea (우리나라에 발생하는 잿빛곰팡이병균 Botrytis cinerea의 분자계통학적 유연관계)

  • Back, Chang-Gi;Lee, Seung-Yeol;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.42 no.2
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    • pp.138-143
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    • 2014
  • Several isolates were collected from apple, pepper, strawberry, cucumber and tomato having typical gray mold symptoms. All the isolates were identified as Botrytis cinerea by using morphological characteristics and PCR-RFLP method. It was difficult to analyze the phylogenetic relationship of these isolates by using ITS region, HSP60 and G3PDH because these genes were highly homologous in their nucleotide in inter-species of B. cinerea and intra-species of genus Botrytis. However, phylogenetic analysis using combined sequences (RPB2, HSP60 and G3PDH genes) clearly showed that all isolate of B. cinerea were different from Botrytis spp. Furthermore, it was also confirmed that strawberry isolate was distantly related to apple, pepper, cucumber and tomato isolates that were closely related to each other in nucleotide level.

Isolation of Brucella spp from sere-positive native bulls and calves below twelve months old (브루셀라병 혈청검사 양성 수소와 12개월령 이하 소에서의 균 분리 및 동정)

  • Ryu, Jae-Yun;Bun, Jung-Hyun;Lee, Hee-Young;Lee, Yong-Chang;Lee, Jong-Jin;Song, Young-Gak;Nam, Hyang-Mi
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.375-384
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    • 2007
  • This study was conducted to investigate the characteristics of brucellosis in Korean native cattle in a farm where bovine brucellosis was confirmed 3 times from September 2006 to March 2007. Of 74 bulls serum samples examined, 21 (28.4%) were positive by Rose-Bengal test (RBT) and Standard tube agglutination test (STAT). In the isolation test from seropositive bulls, B abortus was isolated and identified from 2 specimens (testis, intestinal lymph node) among 6 kinds of specimens including blood, urine, feces and soil. Isolation rate of intestinal lymph node and testis was 25% (3/12 cases) and 16.7% (2/12), respectively. B abortus was also isolated from calves below 12 months old, i.e., 1 isolate (25.0%) was confirmed from testis, 4 (40.0%) from supra-mammary lymph nodes and 1 (25.0%) from intestinal lymph node. All isolates had Brucella specific 16s r-RNA with 905-bp band detected by PCR assay. For the more effective control of bovine brucellosis in korea, this paper would like to suggest that all of bulls and calves should be included in the screening tests.

Characterization of the Stigma side Self-incompatibility Genes in a Self-compatible Brassica oleracea (자가화합성 양배추의 주두측 자가불화합성 유전자 해석)

  • Park, Jong-In;Lee, In-Ho;Jung, Gun-Ho;Nou, Ill-Sup
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1666-1671
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    • 2009
  • In Brassica, S locus glycoprotein (SLG) and S locus receptor kinase (SRK) genes function together for self-recognition in the self-incompatibility response. In addition, a water channel called aquaporins (MOD) is required for the self-incompatibility response. In this study, we isolated the SC-SLG, SC-SRK, and SC-MOD genes from a self-compatible line of B. oleracea. In the self-compatible line, the SC-SLG, SC-SRK, and SC-MOD genes showed the highest degree of sequence similarity with published data and to normal expression by RT-PCR. Therefore, it can be concluded that the SCR/SP11 gene of the B. oleracea pollen may not function and/or that mutations may occur in genes for self-incompatibility that are not linked to the S locus region.

Studies on the Genetic Diversity using RAPD in Leuconostoc sp. Isolated from Kimchi (김치에서 분리한 Leuconostoc속의 RAPD 법을 이용한 유전적 변이도 추론)

  • Kim, Eun-Kyung;Ryu, Chun-Sun;So, Myung-Hwan;Kim, Young-Bae
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.1063-1066
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    • 1997
  • Thirty four Leuconostoc strains originated from kimchi were subjected to analyse their genetical similarity by using RAPD-PCR. At the 0.5 level of similarity, L. mesenteroides subsp. mesenteroides and L. mesenteroides subsp. dextranicum. formed 3 and 2 clusters, and each L. lactis and L. paramesenteroides strain was clustered in one, respectively. Every type strain tested as a control, belongs to the cluster or one of the clusters of the corresponding species or subspecies as the isolates from kimchi, respectively. However the similarity between a and b cluster was less than 0.2 and c cluster is even less similar to both of them (a and b). L. mesenteroides subsp. mesenteroides strains from kimchi have shown a varaiety of RAPD patterns in spite of morphological and physiological similarities.

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First Isolation of Segniliparus rugosus from a Patient with Radiologic Features Similar to Non-Tuberculous Mycobacteriosis (비결핵성 항산균증과 유사한 방사선학적 소견을 보이는 환자에서 Segniliparus rugosus가 동정된 첫 증례)

  • Choi, Sun-Mi;Kang, Hyo-Jae;Jeong, Yun-Jeong;Lim, Joo-Hyun;Choe, Won-Seok;Hwang, Sang-Hyun;Park, Jin-Kyeong;HwangBo, Bin;Lee, Hee-Seok
    • Tuberculosis and Respiratory Diseases
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    • v.72 no.1
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    • pp.82-87
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    • 2012
  • In 2005, a group of mycolic acid-containing bacteria was characterized as belonging to a novel genus, Segniliparus with species Segniliparus rugosus and S. rotundus. We report a case of the S. rugosus isolated from a 54-year-old woman with radiologic features mimicking that of non-tuberculous mycobacteriosis (NTM). When the patient first visited our hospital, an acid-fast bacteria (AFB) smear tested positive and Mycobacterium tuberculosis polymerase chain reaction (TB PCR) was negative in the bronchoalveolar lavage sample. After 2 months, the growing colonies were reported as NTM, but could not be identified because they had died. One year after the initial visit, induced sputum samples showed the same results, positive AFB smear and negative TB PCR. At this point, the growing colonies were identified as S. rugosus. Therefore, we should consider Segniliparus genus as a differential diagnosis for AFB in respiratory specimens in addition to the genus Mycobacterium.

Cloning, characterization, and expression of the macrophage migration inhibitory factor gene from the pacific abalone (Haliotis discus hannai) (북방전복 (Haliotis discus hannai) 의 대식세포이동저해인자 (MIF, Macrophage migration inhibitory factor) 유전자 동정 및 발현분석)

  • Park, Eun Hee;Shin, Eun-Ha;Kim, Young-Ok;Kim, Dong-Gyun;Kong, HeeJeong;Kim, Woo-Jin;An, Chul Min;Nam, Bo-Hye
    • The Korean Journal of Malacology
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    • v.32 no.4
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    • pp.241-247
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    • 2016
  • Macrophage Migration Inhibitory Factor (MIF) are well-defined role as unique cytokine and critical mediator in acute and chronic inflammatory diseases, autoimmune diseases. In this study, we isolated and characterized a full-length of MIF cDNA from the abalone (Haliotis discus hannai). The full-length cDNA of abMIF was of 1264 bp, consisting of a 5'-terminal UTR of 143 bp, an open reading frame of 360 bp and a 3-terminal UTR of 761 bp. The abalone MIF cDNA encodes a 119-amino acid polypeptide with a calculated molecular mass of 13.4 kDa and isoelectric point of 9.07. Multiple alignments and phylogenetic analysis with the deduced abalone MIF protein and showed strong homology with disk abalone (Haliotis discusdiscus). The deduced amino acid sequence of abMIF exhibited homology with other reported MIFs, such as 80%, with that of other disk abalone H. discus discus MIF gene. Quantitative real-time PCR (qRT-PCR) analysis indicated that abMIF was highly expression observed in hapatopacreas, intestine, foot, and gonad of normal conditioned abalone. Even though AbMIF mRNA level in hemocytes was low under the normal condition, it was sharply up-regulated and reached the maximum at 6 h post-infection with Vibrio parahaemolyticus, and then decreased at 24 h post-infection. This result indicates that abMIF plays an important role in responding in the innate immune system.

Isolation of Tetracycline-resistant Lactic Acid Bacteria from Kimchi (김치에서 tetracycline 내성 유산균의 분리)

  • Kang, Hyo-Jin;Kim, Byung-Chun;Park, Wan
    • Korean Journal of Microbiology
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    • v.40 no.1
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    • pp.1-6
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    • 2004
  • Tetracycline resistant bacterial strains were isolated from 10 batches of Kimchi among 50 batches collected in Taegu restrict. The MIC of tetracycline ranged between 25 and> 100 ㎖/l. Total genomic DNA preparation from all 10 tetracycline resistant lactic acid bacterial isolates were subjected to PCR amplification with class-specific primers for tet(M) and tet(O). In only one isolate, HJ9, tet(M) was detected. By Southern blotting and hybridization with a tet(M)-specific probe, the tet(M) gene of HJ9 isolate could be localized on a plasmid. The partial nucleotide sequence and deduced amino acid sequence of tet(M) of HJ9 showed 90-99% and 94-100% homology to those of Gram positive bacteria, respectively. With sequencing of 16S rRNA, HJ9 isolate from Kimchi was identified as Lactobacillus sakei. From these results, Kimchi can be considered potential vehicle for the spread of antibiotic-resistant lactic acid bacteria along the food chain to the consumer.

Detection of viral hemorrhagic septicemia virus (VHSV) in wild marine fishes in the coastal region of Korea (우리 나라 연근해 자연산 해수 어종에서의 Viral Hemorrhagic Septicemia Virus (VHSV)의 검출)

  • Kim, Su-Mi;Park, Su-Il
    • Journal of fish pathology
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    • v.17 no.1
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    • pp.1-10
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    • 2004
  • In order to analyse the detection of viral hemorrhagic septicemia virus (VHSV) in marine environment surrounding coastal region of Eastern and Southern sea of Korea, the pools of each organ sample of three fish were taken for virus assay from February to May in 2003. The samples comprised 42, taken from 9 species of marine fishes. The VHSV was detected from chub mackerel Scomber japonicus and striped mullet Mugil cephalus in epithelioma papulosum cyprini (EPC) cells. The identity of the virus was confirmed a reverse transcriptase-polymerase chain reaction (RT-PCR). VHSV has previously been reported from chub mackerel, but not from striped mullet. The new isolates was classified as a member of genogroup I (American type) of VHSV and was closely related to the VHSV KVHS'01-l based on comparisons of the partial nucleotide sequence of the glycoprotein (G) gene.

Molecular Identification of Thrips in Two Medicinal Crops, Cnidium officinale Makino and Ligusticum chuanxiong Hort (일천궁과 토천궁에서 발생하는 총채벌레류의 분자동정)

  • Jung, Chung Ryul;Jeong, Dae Hui;Park, Hong Woo;Kim, Hyun Jun;Jeon, Kwon Seok;Yoon, Jung Beom
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.1
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    • pp.17-23
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    • 2019
  • Background: Cnidium officinale Makino and Ligusticum chuanxiong Hort. are important medicinal crops in Korea. However, there is insufficient information on the identification of thrips, which attack these plants. Until now, one species of thrips has been recorded as a main pest. Methods and Results: To identify the thrips emerging in C. officinale Makino and L. chuanxiong Hort., these plants were independently cultivated in two local areas. Thirty individuals of each plant species were selected randomly and surveyed for the presence of thrips. After confirming the existence of thrips, 100 thrips individuals were collected from each crop using the beating method. To identify thrips species, we performed PCR-restriction fragment length polymorphism (RFLP)-based analysis using ITS2 primer sets. Six thrips species were identified: western flower thrips (Frankliniella occidentalis), flower thrips (F. intonsa), onion thrips (Thrips tabaci), chrysanthemum thrips (T. nigropilosus), chilli thrips (Scirtothrips dorsalis), and grass thrips (Anaphothrips obscurus). The proportion of these species differed between the host plant species. Conclusions: Six thrips species were major pests of two medicinal crops. Integrated pest management is required to control these thrips species, and will enhance the yield and quality of C. officinale and L. chuanxiong.