• Title/Summary/Keyword: PCR 동정

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The Identification of Alpha-Tubulin as a Biomarker for Diazinon Exposure in Medaka Fish (송사리 모델계에서 다이아지논 노출에 대한 생물 지표로서 알파 튜블린의 동정)

  • Kim, Woo-Keun;Lee, Sung-Kyu;Chon, Tae-Soo;Koh, Sung-Cheol;Kim, Jong-Sang
    • Environmental Analysis Health and Toxicology
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    • v.22 no.4
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    • pp.305-312
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    • 2007
  • 환경오염을 신속하게 모니터링하기 위한 생물지표의 개발은 증가하고 있는 오염의 심각성에 비추어 매우 중요한 과제로 여겨지고 있다. 본 연구에서는 독성물질처리에 의하여 선택적으로 발현이 조절되는 단백질의 동정을 통하여 독성물질에 대한 단백질 생물지표를 발굴하고자 시도하였다. 즉, 송사리(Oryzias latipes)를 유기인계 살충제인 다이아지논(diazinon)에 0, 0.1, 1, 5 mg/L 농도로 24시간 노출시킨 후, 머리와 몸통부분으로 나누어 단백질 발현패턴을 분석하였다. 본 시스템에서 다이아지논 처리에 의하여 유의적으로 발현이 증가된 단백질로서 alpha-tubulin, ribonuclease pancreatic precursor, protein hfq 등을 동정하였으며, 이 가운데 alpha-tubulin과 $hsp90{\beta}$의 발현이 다이아지논 농도에 의존적으로 증가하는 것을 semi-quantitative RT-PCR방법으로 확인하였다. 이와 같이 다이아지논 처리에 특이적으로 발현이 증가된 송사리 단백질들은 노출평가를 위한 생물지표로서 개발에 응용될 수 있을 것으로 평가된다.

Domestic Unrecorded Species Found in the Eastern Part of Jeju Island (제주 동부지역에서 발견된 국내 미기록 자생버섯 보고)

  • Lee, Seung Hak;Ko, Pyung Yeol;Jeun, Yong Chull
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.105-113
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    • 2018
  • In this study, we report the occurrence of four domestic mushrooms (among 317 samples) that were previously undocumented in the eastern part of Jeju Island in Korea. To identify the mushrooms, DNA sequences of the mushrooms were analyzed by comparing them with the data available in NCBI GenBank. Mushrooms were identified as Agaricus flocculosipes, Amanita concentrica, Coprinopsis strossmayeri, and Favolus acervatus, none of which have been previously documented in Korea. Furthermore, microscopic observations revealed that the morphology of the four mushrooms was consistent with the features of the species matched by the DNA sequences analysis.

수계 생태계에서의 세균 군집 구조의 분자생물학적 분석

  • 이동훈;김상종
    • Korean Journal of Microbiology
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    • v.33 no.1
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    • pp.55-65
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    • 1997
  • 16S rRNA를 분석한 연구들은 자연 생태계에서 추출한 핵산을 이용하여 16rRNA 유전자의 염기서열을 분석하거나 특정 DNA probe를 이용한 hybridization 실험이 주류를 이루어 왔다. 특히 PCR 기법이 개발됨에 따라 적은 양의 시료를 대량으로 손쉽게 증폭시킬 수 있어 다양한 분야에 응용되고 있다. 세균 군집의 구조를 이해하는데 있어서 PCR 방법의 적용 대상은 주로 16S rRNA 유전자의 염기서열 해독분야이며 해양 생태계를 대상으로 많은 연구 결과가 보고되었다(11,13,21,26). 한편 자연 생태계의 개별적 미생물 분류룬들을 검출하기 위한 특정 oligonucleotide probe의 개발방법들은 미생물 군집의 유전적 다양성에 대한 정보 파악 이외에 배양이 어려운 혐기성 세균과 같은 특정 세균들의 동정에도 이용되고 있다(3,24,55). 본고에서는 세균 군집의 구조와 다양성을 연구하는데 적용 가능한 rRNA 분석방법들을 수계 생태계를 중심으로 살펴보고자 한다.

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Characterization of Tetracycline Resistance Plasmids of Staphylococcus aureus (황색포도상구균의 테트라사이클린 내성 플라스미드 동정)

  • Park, Jung-Hee;Lee, Jae-Yoon;Moon, Kyung-Ho
    • YAKHAK HOEJI
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    • v.52 no.4
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    • pp.279-282
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    • 2008
  • Plasmids were isolated from 15 tetracycline (Tc) resistant S. aureus. Two small tetracycline resistance plasmids, pKH16 and pKH17, have been isolated from Staphylococcus aureus JY10 and Staphylococcus aureus JY22, respectively and the complete nucleotide sequences of those plasmids have been determined. pKH16 consisted of 4,442 bp and showed high identity to pKH6 (99% matching percentage) isolated in 1989 from S. aureus SA2. pKH17 consisted of 4,441 bp and showed less identity to pKH6 (95% matching percentage) than pKH16. PCR analysis showed that tetK and tetM did not exist in ten large plasmids isolated from ten Tc resistant S. aureus. Twelve Tc resistant S. aureus showed reistance both to Tc and Mn and we might analogize that twelve Tc resistant S. aureus had tetM in their chromosome.

Isolation of labial Gene in the Oligochaeta Perionyx excavatus and Expression Patterns during Head Regeneration (지렁이 labial 유전자의 동정 및 머리재생 시 발현 양상에 관한 연구)

  • Cho Sung Jin;Lee Myung Sik;Tak Eun Sik;Hur So Young;Lee Jong Ae;Park Bum Joon;Cho Hyun Ju;Shin Chuog;Park Soon Cheol
    • The Korean Journal of Soil Zoology
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    • v.8 no.1_2
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    • pp.1-6
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    • 2003
  • Hox genes are pivotal in the control of morphogenesis along the anterior-posterior (AP) axis in various bilaterians. Several indications suggest their involvement in the control of cell growth and regeneration. For the labial full-length fragment, RACE-PCR was employed to obtain the 3' and 5' franking regions. Semi-quantitative RT-PCR analysis revealed that the labial expression began to increase at 12 hours after amputation. The peak expression was approximately 1.5-fold more than the unamputated controls. This result could give us information on the significance of Hox genes and the relationships between Hox genes during regeneration.

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PCR-based Identification of Eubacteirum species in endodontic infection (감염 근관에서 중합효소연쇄반응법을 이용한 Eubacterium 균종의 동정)

  • Kum, Kee-Yeon;Fouad, A.F.
    • Restorative Dentistry and Endodontics
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    • v.28 no.3
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    • pp.241-248
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    • 2003
  • Asaccharolytic Eubacterium균종은 감염 근관에서의 높은 발생 빈도와 독성으로 인해 최근 많이 연구되어지고 있다. 본 연구는 24명의 환자의 감염 근관에서 얻은 22개의 PCR 산물로부터 Eggerthella lenta를 포함한 Eubacterium 균종의 빈도 및 환자의 임상 증상이나 당뇨와의 상관성을 조사한 후 얻은 자료를 토대로 다음과 같은 결과를 얻었다. 1. 22개의 표본 중에서 16개(73%)가 Eubacterium균종을 포함하고 있었으며 이 중 9개의 시편에서 Eubacterium infirmum이 검출되었다. 2. Eggerthella lenta는 어떤 시편에서도 발견되지 않았다. 3, Odds ratio analysis 결과 Eubacterium infirmum은 당뇨병과의 높은 상관성을 보여주었다(OR=9.6, P=0.04).

Detection of cry-type Genes of Bacillus thuringiensis Isolates from Korea (한국에서 분리한 Bacillus thuringiensis 균주의 cry형 유전자의 동정)

  • Park Sooil;Lee Kwang Yong;Kang Eun Young;Kim Eui Na;Kwon Hyuk Han;Ahn Seong Kyu;Lee Hyung Hoan
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.154-158
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    • 2005
  • Twenty-three Bacillus thuringiensis strains isolated from Korea were screened to detect the cry-type genes using PCR with 21 specific oligonucleotide primers. Eight strains contained distinct multiple crystal genes; cry1Aa2, cry1Ab1, cry1Ac1 and cry2Aa1. These results indicate that the strains coincided with the B. thuringiensis subsp. kurstaki strain. The other 15 strains were not recognised to the 21 specific primers.

Molecular identification of Anisakid worm third stage larvae isolated from masou salmon Oncorhynchus masou (시마연어, Oncorhynchus masou에서 분리된 아니사키스 속 선충 3기 유충의 분자생물학적 방법을 이용한 동정)

  • Jeon, Chan-Hyeok;Setyobudi, Eko;Kim, Jeong-Ho
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.421-427
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    • 2010
  • Anisakid nematodes third stage larvae were isolated from the muscles of masou salmon (Oncorhynchus masou). Fish were purchased from Jumunjin fishery market in Gangneung. Four Anisakid third stage larvae were isolated from 4 fish. Molecular identification of the isolated worms was conducted by PCR-RFLP analysis of ribosomal DNA internal transcribed spacer region and direct sequencing of mitochondrial DNA cox2 gene. As results, all the tested individual worms were identified as Anisakis simplex (sensu stricto). This is the first report of molecular detection of anisakid worms in salmonid fishes in Korea.

Isolation of Myxobacteria Carrying Soraphen Biosynthetic Gene Clusters (Soraphen 생합성 유전자군을 갖는 점액세균의 분리)

  • Lee, Cha-Yul;Hyun, Hye-Sook;Cho, Kung-Yun
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.10-16
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    • 2009
  • Polymerase chain reactin (PCR) could be a simple way to screen new microbial strains producing useful secondary metabolites if their biosynthetic genes are known and candidate strains to be screened are available. In this study, we have screened two myxobacterial strains, KYC3047 and KYC3076, carrying genes appeared to be biosynthetic genes of soraphen A, a potent antifungal substance, out of 50 cellulose degrading myxobacteria using PCR. The two strains were identified as Sorangium cellulosum based on morphological, physiological, and molecular biological characteristics. Both of the strains produced substances having strong antifungal activities as expected against Candida albicans, a causative agent of candidiasis, and Colletotrichum acutatum, a causative agent of anthracnose on pepper.

First Report of Cucumber mosaic virus Isolated from Sambungai (Gynura procumbens) (명월초에서 분리한 오이모자이크바이러스의 감염 첫 보고)

  • Kwon, June;Hong, Jin-Sung
    • Research in Plant Disease
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    • v.23 no.4
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    • pp.379-382
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    • 2017
  • In March 2016, an isolate of Cucumber mosaic virus (named Gyp-CMV) was isolated from the Sambungai (Gynura procumbens) showing the symptoms of mosaic and chlorosis. The isolate Gyp-CMV was characterized by disease reactions in several indicator plants, reverse transcription-polymerase chain reaction (RT-PCR), PCR-restriction fragment length polymorphism, and sequence analysis of movement protein (3a) and coat protein (CP) genes. Tobacco, tomato, pepper, ground cherry, and lambsquarters (Chenopodium quinoa and C. amaranticolor) appeared typical CMV symptoms, but zucchini and cucumber were not infected. Phylogenetic analysis of the 3a and CP gene indicated that Gyp-CMV belongs to the CMV subgroup II. Sequence identities of the Gyp-CMV 3a and CP genes showed 99.3% and 100% to that of Hnt-CMV at amino acid level. To our knowledge, this is the first report of CMV infection in Gynura procumbens.