• 제목/요약/키워드: Internal transcribed spacer(ITS)

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ITS-RFLP와 ITS1 염기서열 분석에 의한 피부사상균의 동정과 계통적 유연관계 (Identification and Phylogenetic Relationship of Dermatophytes Based on RFLP Analysis and Nucleotide Sequence of Internal Transcribed Spacer (ITS)1 in Nuclear Ribosome DNA)

  • 최연화;이영선;유재일;김봉수
    • 대한미생물학회지
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    • 제35권1호
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    • pp.49-60
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    • 2000
  • ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp..

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Biomass and Molecular Characteristics of Multi-tillering Miscanthus Mutants

  • Lee, Geung-Joo;Zhang, Lili;Choi, Young In;Chung, Sung Jin;Yoo, Yong Kweon;Kim, Dong Sub;Kim, Sang Hoon
    • 한국자원식물학회지
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    • 제25권6호
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    • pp.745-752
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    • 2012
  • Compared to wide ranges of genetic variation of natural populations, very limited Miscanthus cultivar has been released. This study was the first report on the development of Miscanthus cultivar by means of radiation breeding. Seeds of M. sinensis were initially exposed to gamma rays of 250 Gy for 24 h, generated from a $^{60}Co$ gamma-irradiator. The irradiated seeds were sown and then the highly tiller-producing mutants were selected for this study. Biomass-related parameters including tiller number, plant height, stem diameter, and leaf number were measured. Ploidy level and internal transcribed spacer (ITS) were investigated to characterize the mutants compared to wild type (WT) Miscanthus. Plant height and tiller number were negatively related, where multi-tillering mutants were relatively short after 4 month growth. However stem diameter and leaf number were greater in mutants. All the materials used in this study were diploid, implying that the mutants with greater tiller numbers and stem diameter were not likely related to polyploidization. Based on the sequence of ITS regions, the mutants demonstrated base changes from the gamma irradiation where G+C content (%) was decreased in the ITS1, but increased in ITS2 when compared to WT sequence. ITS2 region was more variable than in ITS1 in the mutants, which collectively allows identification of the mutants from WT. Those mutants having enhanced tillers and allelic variations might be used as breeding materials for enhanced biomass-producing Miscanthus cultivars.

개와 고양이 유래 피부사상균의 분자생물학적 계통 분석 (Molecular Phylogenetic Classification of Dermatophytes Isolated from Dogs and Cats)

  • 김두;정석영;안소저
    • 한국임상수의학회지
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    • 제23권4권
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    • pp.405-410
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    • 2006
  • 피부사상균증이 있는 개와 고양이에서 분리한 9주의 Microsporum canis와 5주의 Microsporum gypseum에서 ribosomal DNA를 추출하여 internal transcribed spacer 1 (ITS1) gene을 PCA로 증폭한 후 sequencing을 실시하여, 각 사상균의 계통학적 관계를 조사하였다. M canis 분리주 9주의 ITS1 gene의 nucleotide sequence는 100% 일치하였으며 M gypseum 분리주 5주의 nucleotide sequence도 100% 일치하였다. M canis 분리주 9주의 계통분석 결과 미국, 일본, 호주 및 유럽에서 분리된 M canis와 같은 cluster에 속하였으며 다른 Microsporum spp와는 유전적으로 다른 cluster를 형성하였다. 그러나 M canis와 M distortum, M equinum, M ferrugineum은 유전적으로 매우 가까운 위치에 있었다. M gypseum 분리주는 M canis와는 다른 cluster를 형성하였다. ITS1 gene의 분자생물학적 분석은 Microsporum spp를 확인하고 그들의 유전학적 관계를 이해하는 유용한 정보를 제공하는 것으로 생각된다.

Use of Restriction Fragment Length Polymorphism Analysis to Differentiate Fungal Strains in Sunchang Meju

  • Jung, Jong-Hyun;Seo, Dong-Ho;Bhoo, Sung-Hee;Ha, Suk-Jin;Kim, Jong-Sang;Kim, Jeong-Hwan;Kwon, Dae-Young;Cha, Jae-Ho;Park, Cheon-Seok
    • Food Science and Biotechnology
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    • 제17권4호
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    • pp.888-891
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    • 2008
  • Twenty-three fungal strains were isolated from meju that had originated from the Sunchang province, the famous location for making fermented soybean foods in Korea. The restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS) region of the rDNA (ITS-RFLP) was applied to differentiate the isolated fungal strains. First, the ITS region by polymerase chain reaction (PCR) with specific primers was amplified and then cleaved the products with different restriction enzymes. Cleavage of the amplified fragments with the restriction enzymes AluI, HaeIII, HhaI, and TaqI revealed extensive polymorphisms. The ITS-RFLP results highly correlated with ITS sequence analysis. All of the 23 fungal strains were classified into 5 groups by ITS-RFLP analysis. Aspergillus oryzae was the major fungal strain isolated from Sunchang meju (12 out of 23), while Aspergillus fumigatus was the next most frequently isolated strain (7 out of 23). In contrast, it was found that Fusarium asiaticum, Aspergillus sydowii, and Arthrinium sp. were the minor fungal strains in meju.

RFLP를 이용한 Gyrodactylus salaris의 internal transcribed spacer(ITS) PCR 위양성 판별 (Determination of false positives in PCR diagnostics based on the internal transcribed spacer (ITS) of Gyrodactylus salaris using RFLP)

  • 김민성;최희정;정지민;권문경;황성돈
    • 한국어병학회지
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    • 제37권1호
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    • pp.147-153
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    • 2024
  • The World Organization for Animal Health (WOAH) recommends two protocols (ITS and COI) for conventional PCR of G. salaris diagnosis. However, ITS PCR protocol may yield false-positive results, leading to unnecessary countermeasures. It's difficult to distinguish between G. salaris and false-positive by similar amplicon size of PCR, since the amplicon size of ITS PCR in G. salaris and false-positive was 1,300 and 1,187 bp, respectively. The nucleotide sequences of ITS false-positive in rainbow trout is 99.7% identical to previously reported host genome sequences of rainbow trout (Oncorhynchus mykiss) and 95.3 to 89.1% identical to those of other salmonid fish species. To reduce false-positive PCR band, PCR was performed by the different annealing temperature, but PCR bands were still detected. In RFLP analysis by HaeIII, the PCR product of G. salaris was digested into four bands of 512, 399, 234 and 154 bp, while the false-positive was digested into seven bands of 297, 263, 242, 144, 93, 80 and 68 bp. In the RFLP patterns digested by HindIII, G. salaris showed two bands of 659 and 640 bp, while false-positive had one fragment of 1,187 bp without any digestion. Therefore, the RFLP method of ITS PCR with HaeIII and HindIII can be used for differentiation between G. salaris and false-positive. These results might provide important information on the improvement of PCR diagnostic method of G. salaris.

Intrageneric Relationships of Trichoderma Based on Internal Transcribed Spacers and 5.8S rDNA Nucleotide Sequences

  • Kim, Gi-Young;Lee, Goang-Jae;Ha, Myung-Gyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • 제28권1호
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    • pp.11-16
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    • 2000
  • The nucleotide sequences of the internal transcribed spacer (ITS) regions of the ribosomal DNA including the 5.8S ribosomal RNA gene (rDNA) have been determined for 11 species in order to analyze their intrageneric relationships. The total length of these sequences ranged from 530 nucleotides for Trichoderma reesei KCTC 1286 to 553 nucleotide for Trichoderma koningii IAM 12534. Generally speaking, the length of ITS1 region was about 30 nucleotides longer than that of the ITS2 region. Also, the sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites were also found. Thus, a neighbor-joining tree was constructed using the full sequence data of the ITS regions and the 5.8S rDNA. The Trichoderma genus used to be grouped on the basis of the morphological features and especially the shape of phialides needs to be reexamined. The phylogenetic tree displayed the presence of monophylogeny in the species of Trichoderma. Therefore, it was difficult to distinguish the intrageneric relationships in the Trichoderma genus.

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백년초선인장의 ITS(internal transcribed spacer) 유전자 분석 (Analysis of the ITS (Internal Transcribed Spacer) Region of Opuntia ficus-indica)

  • 인준교;이범수;김은정;최관삼;한승호;신철우;양덕춘
    • 한국자원식물학회지
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    • 제19권1호
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    • pp.161-168
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    • 2006
  • 제주도에 자생하는 부채 선인장인 백년초의 기원 규명을 목적으로 ITS primer를 이용하여 685 bp의 ITS 영역을 분리하였다. ITS 영역의 염기서열을 분석한 결과 18S rRNA의 길이는 54 bp, 26S rRNA는 55 bp, ITS1은 193 bp, ITS2는 220 bp로 구성되어 있었다. 백년초 ITS 영역은 기존에 보고된 Cucurbitoideae 식물들의 ITS 영역에 비하여 ITS2 스페이서 영역의 239-254 bp보다는 다소 짧았다. 그러나 이들 스페이서 영역의 GC 함량은 백년초의 경우 ITS1은 66.8%, ITS2의 경우에는 67.7%로 Cucurbitoideae 식물들에서 보다 높은 GC 함량을 나타내었다. 백년초 선인장의 rDNA 영역에 가장 높은 상동성을 나타낸 것은 같은 Opuntioideae에 속하는 Pereskiopsis porteri(L78037)로 95%의 유사도를 나타내었다. 백년초 rDNA Clustal W 프로그램을 이용하여 유연관계를 조사한 결과 같은 Opuntioideae에 속하는 Pereskiopsis porteri(L78037)와 같은 cluster로 분리되었다.

Identification of Entomopathogenic Fungus, Beauveria spp. F-101 Isolated from Thecodiplosis japonensis Using Internal Transcribed Spacer Sequence

  • Shin, Sang-Chul;Roh, Jong-Yul;Kim, Chul-Su;Park, Il-Kwon;Jeon, Mun-Jang;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • 제8권1호
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    • pp.77-80
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    • 2004
  • For the development of the alternative control system against the major forest pests, Beauveria spp. F-101, isolated from a dead larva of Thecodiplosis japonensis, was selected because this isolate showed high pathogenicities against T. japonensis and Acantholyda parki. Beauveria spp. F-101 had irregular clustered conidio-phores and conidia borne on a distinctive apical zigzag extension, and it showed typical characteristic of the genus, Beauveria in morphology. For molecular based-identification, the ribosomal ITS region of Beauveria spp. F-101 was amplified with ITS1 and ITS4 primers, and cloned into pGEM- T Easy vector. The amplified PCR product was 569 bp in size and completely sequenced. The similarities of the cloned ITS sequence were 99 % and 97% to those of B. bassiana and B. brongniartii, respectively. In comparison to other species among the genus Beauveria, the ITS region of Beauveria spp. F-101 showed a similarity of 95% to B. amorpha, 95% to B. tenella, 89% to B. vermiconia and 69% to B. alba, respectively. In addition, in comparison to different genus, it had 95% similarities to Cordyceps militaris and 91% to Paecilomyces tenuipes. Accordingly, the current result suggests that Beauveria spp. F-101 was a variant of B. bassiana and it seems to be a new isolate considering sequence variation in ITS region.

고려인삼과 미국삼의 종간잡종으로부터 재분화된 식물체의 특성 (Characteristics of Plantlets Redifferentiated from F1 Hybrid between Panax ginseng and Panax quinquefolius)

  • 안인옥;이성식;이장호;이범수;인준교;양덕춘
    • Journal of Plant Biotechnology
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    • 제33권1호
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    • pp.45-48
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    • 2006
  • 교잡 1세대의 화서조직에서 재분화된 F1 유식물체는 고려인삼 유식물체에 비하여 지상부와 지하부 생육이 모두 양호하였으며, 줄기 색도 고려인삼 재분화 식물체의 줄기에 비하여 자색을 강하게 띄었으며 잎의 색도 진한 녹색을 나타내었다. 천풍, 연풍, 선원 등의 고려인삼의 품종 내에서는 Internal Transcribed Spacer (ITS)영역의 DNA PCR 패턴간에 차이점이 나타나지 않았으나, 고려인삼과 미국인삼은 각기 다른 PCR 패턴을 보였으며, 고려인삼과 미국인삼간의 교잡 1세대는 고려인삼과 미국인삼에 나타나는 고유한 PCR패턴을 모두 나타내었다. 교잡 1세대에서 유기한 캘러스와 재분화식물체는 조직배양 모본인 교잡 1세대와 동일한 PCR 패턴을 보임에 따라 교잡 1세대의 조직배양체는 ribosomal DNA의 ITS영역에서 유전적인 안정성을 나타내는 것으로 확인되었다.