• 제목/요약/키워드: rDNA ITS region

검색결과 251건 처리시간 0.036초

Development of Rapid Molecular Detection Marker for Colletotrichum spp. in Leaf and Fruit Tissues of Sweet Persimmon

  • Iee, Sang-Pyo;Lee, Youn-Su
    • Journal of Microbiology and Biotechnology
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    • 제12권6호
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    • pp.989-992
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    • 2002
  • Sweet persimmon (Diospyros kaki Thunb.) is widely cultivated in the southern part of Korea and its cultivation is increasing. However, anthracnose disease caused by Colletotricuhum species is one of the major hinderances to the cultivation and production of sweet persimmon. Therefore, in the current study, PCR was used to specifically detect Colletotrichum spp., based on the sequences of the ITS II regions in the rDNA. Using the sequence data, CO-1 was designated to detect Colletotrichum together the with ITS 4 primer. The result showed that a single segment of ca. 500 bp was observed only in Colletotrichum, but not in any other fungal and bacterial isolates. The annealing temperatures and template DNA quantites were also investigated to identify optimal conditions for detection. Using these species-specific primers, a unique band was obtained at annealing temperatures ranging from $55^{\circ}C\;and\;61^{\circ}C$ and template DNA levels from 10 pg- $10{\mu}g$.

rDNA-ITS DNA 바코드 부위 분석을 통한 산초(山椒) 기원종 감별용 유전자 마커 개발 (Development of Molecular Markers for the authentication of Zanthoxyli Pericarpium by the analysis of rDNA-ITS DNA barcode regions)

  • 김욱진;지윤의;이영미;강영민;최고야;문병철
    • 대한본초학회지
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    • 제30권3호
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    • pp.41-47
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    • 2015
  • Objectives : Due to the morphological similarity of the pericarp and description of multi-species in National Pharmacopoeia of Korea and China, the Zanthoxylum Pericarpium is difficult to authenticate adulterant in species levels. Therefore, we introduced the sequence analysis of DNA barcode and identification of single nucleotide polymorphism(SNP) to establish a reliable tool for the distinction of Zanthoxylum Pericarpium from its adulterants. Methods : To analyze DNA barcode region, genomic DNA was extracted from twenty-four specimens of authentic Zanthoxylum species and inauthentic adulterant and the individual internal transcribed spacer regions (rDNA-ITS and ITS2) of nuclear ribosomal RNA gene were amplified using ITS1, ITS2-S2F, and ITS4 primer. For identification of species-specific sequences, a comparative analysis was performed using entire DNA barcode sequences. Results : In comparison of four Zanthoxylum ITS2 sequences, we identified 16, 4, 6, and 4 distinct species-specific nucleotides enough to distinguish Z. schinifolium, Z. bungeanum, Z. piperitum, and Z. simulans, respectively. The sequence differences were available genetic marker to discriminate four species. Futhermore, phylogenetic relationship revealed a clear classification between different Zanthoxylum species showing 4 different clusters. These results indicated that comparative analysis of ITS2 DNA barcode was an useful genetic marker to authenticate Zanthoxylum Pericarpium in species levels. Conclusions : The marker nucleotides, enough to distinguish Z. schinifolium, Z. piperitum, Z. bungeanum, and Z. simulans, were obtained at 30 SNP marker nucleotides from ITS2 sequences. These differences could be used to authenticate official Zanthoxylum Pericarpium from its adulterants as well as discriminating each four species.

한국산 방사무의김 (Porphyra yezoensis)의 핵 18S rDNA 염기서열 분석 (Sequence Analysis of Nuclear 18S rDNA from the Seaweed Porphyra yezoensis (Rhodophyta) in Korea)

  • ;김명숙;최재석;조지영;진형주;홍용기
    • 한국수산과학회지
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    • 제35권6호
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    • pp.633-638
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    • 2002
  • Nuclear 18S ribosomal RNA gene (185 rDNA) from the aquaculturable seaweed Porphya yezoensis (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analysed. Complete 185 rDNA has an 1823 bp exon and a 514 bp intron. The G+ C contents of exon and intron were $48\%$ and $51.4\%$, respectively. The exon sequence showed $99.5\%$ homology to the GenBank accession number AB013177 of the Japanese p. yezoensis. The intron region that was inserted upstream between 568 and 1083 showed $93.4\%$ homology to the AB013177.

Sequence Analysis of the Internal Transcribed Spacer of Ribosomal DNA in the Genus Rhizopus

  • Park, You-Jung;Min, Byung-Re
    • Mycobiology
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    • 제33권2호
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    • pp.109-112
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    • 2005
  • The internal transcribed spacer (ITS) regions including the 3'-end of 18S rRNA gene, 5.8S rRNA gene and the 5'-end of the 28S rRNA gene of Rhizopus spp. were amplified by PCR and analyzed by DNASIS program. Length polymorphism of these region ranged from 564 bp in R. oryzae to 789bp in R. stolonifer. The length and sequence of 5.8S was very conserved with $154{\sim}155\;bp$. The sequence of ITS2 was more variable than that of ITS1. The base substitution rates were ranged from 0 to 0.6069 per site, and higher rate was found in R. stolonifer. In general, transition was usually more frequent than transversion. On the basis of sequencing results, four groups were clustered with value of 61.9% similarity; R. oryzae, R. micros pores, R. homothallicus, and R. stolonifer groups.

Vegetative Growth and Phylogenetic Relationship of Commercially Cultivated Strains of Pleurotus eryngii based on ITS sequence and RAPD

  • Alam, Nuhu;Shim, Mi-Ja;Lee, Min-Woong;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, Tae-Soo
    • Mycobiology
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    • 제37권4호
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    • pp.258-266
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    • 2009
  • Pleurotus eryngii, known as king oyster mushroom has been widely used for nutritional and medicinal purposes. This study was initiated to screen the suitable conditions for mycelial growth and to determine the phylogenetic relationship of the selected strains. Optimal mycelial growth was observed at $30{^{\circ}C}$ and minimum mycelial growth observed at $10{^{\circ}C}$. This mushroom tolerates a broad pH range for mycelial growth, with most favorable growth observed at pH 6. Results also indicated that glucose peptone, yeast malt extract and mushroom complete media were favorable growth media, while Hennerberg and Hoppkins media were unfavorable. Dextrin was the best and xylose the least effective carbon sources. Results revealed that inorganic nitrogen sources were less effective than organic sources for the mycelial growth of P. eryngii. Investigation of genetic diversity is necessary to identify the strains. The ITS region of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 214 to 222 bp and 145 to 236 bp, respectively. The sequence of ITS2 was more variable than that of ITS1, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into six clusters. Fourteen IUM and ATCC- 90212 strains were also analyzed by RAPD with 20 arbitrary primers. Fourteen of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.2 to 2.3 kb.

국내 미기록종 선충포식성 곰팡이 Drechslerella brochopaga Kan-23의 형태 및 계통분류 (Morphological and Phylogenetic Characteristics of a Nematophagous Fungus, Drechslerella brochopaga Kan-23)

  • 조천휘;강두선;김윤지;황경숙
    • 미생물학회지
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    • 제44권1호
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    • pp.63-68
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    • 2008
  • 참외 경작지 토양으로부터 순수 분리된 선충포식성 곰팡이 Kan-23 균주는 배양 30일 이후에 분생포자가 형성되는 증식이 매우 더딘 특성을 나타내었다. 주사전자현미경(SEM)을 이용하여 Kan-23 균주의 미세형태 구조를 관찰한 결과 분생포자병의 각 가지마다 $5{\sim}10$개의 분생포자를 형성하였으며, 분생포자는 3개의 격막을 형성하고 타원형을 나타내었고, 수축성 고리형태(constricting ring)의 포식 구조체를 형성하는 특징을 나타내었다. 다양한 배지를 이용하여 포식 구조체의 형태학적 특성을 비교 검토한 결과 GPA 배지에서 2배 이상 크기의 대형 수축성 고리 형태의 포식 구조체를 형성하는 특징을 나타내었다. Kan-23 균주의 rDNA의 ITS 영역의 염기서열을 분석한 결과, Drechslerella 속의 계통군에 속하였으며, 특히 Drechslerella brochopaga (U51950)와 99%의 높은 염기서열 유사도를 나타내어 Drechslerella brochopaga로 동정되어 국내 미기록종임이 확인되었다.

Promoter demethylation mediates the expression of ZNF645, a novel cancer/testis gene

  • Bai, Gang;Liu, Yunqiang;Zhang, Hao;Su, Dan;Tao, Dachang;Yang, Yuan;Ma, Yongxin;Zhang, Sizhong
    • BMB Reports
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    • 제43권6호
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    • pp.400-406
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    • 2010
  • Cancer/testis (CT) antigens exhibit highly tissue-restricted expression and are considered promising targets for cancer vaccines. Here we identified a novel CT gene ZNF645 which restrictively expresses in normal human testes and lung cancer patients (68.3%). To investigate the promoter methylation status of ZNF645, we carried out bisulfite genomic sequencing and found that the CpG island in its promoter was heavily methylated in normal lung tissues without the expression of ZNF645, whereas there was high demethylation in normal human testes and lung carcinoma tissues with its expression. Also ZNF645 could be remarkably activated in A549 and HEK293T cells treated by DNA demethylation agent 5'-aza-2'-deoxycytidine. And the dual luciferase assay revealed that the promoter activity of the ZNF645 was inhibited by methylation of the CpG island region. Therefore, we proposed that ZNF645 is a CT gene and activated in human testis and lung cancers by demethylation of its promoter region.

청미래덩굴의 근권에서 분리된 2종의 Glomeromycota 미기록종 (Two Unreported Glomeromycota Fungi Isolated from Rhizospheres of Smilax china)

  • 박혁;가강현;엄안흠
    • 한국균학회지
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    • 제47권3호
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    • pp.275-280
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    • 2019
  • 청미래덩굴의 근권 토양을 온실에서 배양하여 Glomeromycota에 속하는 균류의 포자를 분리하였다. 분리된 포자는 18S partial rDNA 영역의 DNA 염기서열 분석과, 더 정확한 동정을 위해 추가적으로 ITS 영역과 28S rDNA 영역까지 포함하는 $Kr{\ddot{u}}ger$ fragment 지역의 염기서열을 분석하여 동정하였다. 동정 과정에서 2종의 국내 미기록종 Glomeromycota 균류의 포자를 확인하였으며, 확인된 종은 Diversispora eburnea, Paraglomus laccatum이다. 확인된 미기록종 포자의 형태적 특성 및 계통적 분석의 결과에 대해 기술하였다.

헐떡이풀의 핵형분석과 Bicolor FISH를 이용한 물리적 지도 작성 (Karyotype Analysis and Physical Mapping of rDNAs Using Bicolor-FISH in Tiarella polyphylla D. Don)

  • 김수영;이중구
    • 한국자원식물학회지
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    • 제20권5호
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    • pp.446-450
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    • 2007
  • 헐떡이풀은 다년생 초본으로 중국, 일본, 대만 그리고 한국에 분포한다. 특히 우리나라에서는 울릉도에서만 분포하는데, 천식 치료, 타박상 그리고 청각장애의 치료에 사용된다. 약용작물로써의 높은 가치에도 불구하고 염색체 수를 제외한 다른 세포유전학적인 연구가 거의 이루어지지 않았다. 따라서 핵형분석 뿐만 아니라 bicolor FISH를 통한 5S 와 45S rDNA의 물리적 지도작성에 관한 연구가 수행되었다. 체세포 염색체 수는 2n=2x=14로 염색체의 길이는 $1.66{\sim}3.50{\mu}m$ 이다. 또한 염색체의 구성은 4쌍의 차중부 염색체(염색체 1, 2, 3, 6)와 2쌍의 차단부 염색체(염색체 5, 7)그리고 1쌍의 단부 염색체(염색체 4)로 확인되었다. 또한 4번 염색체가 부수체 염색체로 관찰되었다. Bicolor-FISH를 통해 각각 1쌍의 5S와 45S rDNA 위치를 확인하였는데, 5S rDNA의 경우 염색체 3번의 동원체 부위에서 확인되었고, 45S rDNA는 염색체 4번의 단완 말단 부위에서 관찰되었다. Bicolor-FISH는 헐떡이풀 염색체상에 rDNA 유전자의 위치 확인에 매우 유용한 정보를 제공하는 기술로 사용되었다.