• Title/Summary/Keyword: ITS rDNA

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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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    • v.43 no.6
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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.

Molecular phylogeny of moon jellyfish Aurelia aurita Linnaeus collected from Yeosu waters in Korea based on nuclear and mitochondrial DNA sequences (여수해역에서 채집한 보름달 둥근 물해파리의 핵과 미토콘드리아 DNA 염기서열을 이용한 유연 관계 분석)

  • Kim, Sook-Yang;Cho, Eun-Seob
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.318-327
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    • 2007
  • This study presents the molecular phylogenetic analysis of Korean Aurelia aurita Linnaeus collected from Yeosu in the southern waters of Korea using nuclear ITS1 region and mitochondrial COI gene sequences. The use of oligonucleotide primers F5 (forward) and R5 (reverse) targeted to ITS1 and LCO1490 (forward) and HCO2198 (reverse) targeted to COI amplified 267 bp and 643 bp fragments, respectively. The shortest genetic distance towards the ITS1 region is estimated at 0.023 when comparing Korean A. aurita to Aurelia sp. collected from California, USA. In particular, Korean and American/Swedish A. aurita were located far away in terms of genetic distance, ranging from 0.393 to 0.395. On the other hand, the genetic distance between Korean and English/Turkish/Swedish/American A. aurita regarding the mitochondrial DNA COI gene ranged from 0.201 to 0.205. However, a sister-ship with Korean and American A. aurita showed an extremely high bootstrap value (100%). The predicted secondary RNA structure of the mitochondrial DNA COI gene showed many different folding structures with a similar energy between Korean and American A. aurita. These results suggest that ITS1 and the mitochondrial DNA COI gene could be used as genetic markers for identification of the biogeographic populations.

Phylogenic Relationships of Rubus Species Revealed by Randomly Amplified Polymorphic DNA Markers

  • Eu, Gee-Suck;Chung, Byung-Yeoup;Bandopadhyay, Rajib;Yoo, Nam-Hee;Choi, Dong-Geun;Yun, Song-Joong
    • Journal of Crop Science and Biotechnology
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    • v.11 no.1
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    • pp.39-44
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    • 2008
  • Korean cultivated bramble, which is known as Bokbunja-ddal-gi is regarded to be originated from Korea native Rubus coreanus. However, little scientific evidence and significant morphological differences between Korean cultivated bramble(KCB) and R. coreanus throw doubt on the ancestry of KCB. This study was carried out to obtain phylogenetic information on KCB by comparing its nuclear genomic background with those of R. coreanus, black(R. occidentalis) and red(R. idaeus) raspberry, blackberry(R. lanciniatus) and R. crataegifolius. A total of 99 random amplified polymorphic DNA(RAPD) markers were generated and used for phylogenetic analysis of 76 Rubus accessions. Accessions of each species were grouped into each distinct subclade by the RAPD markers at a similarity coefficient of about 0.59. The KCB subclade formed a clade with R. occidentalis and R. crataegifolius subclades at a similarity coefficient of 0.47. The R. coreanus subclade formed a clade with R. idaeus, R. lanciniatus and R. crataegifolius subclades at a similar similarity coefficient. Only one KCB accession from Hoengsung was included in R. coreanus subclade. The accession shows leaf and flower characteristics different from the rest of the KCB accessions. The phylogenetic relationship inferred from the RAPD markers suggests that the nuclear genomic background of KCB accessions which show morphological similarity to black raspberry is more closely related to black raspberry than to R. coreanus. This brings about the need for close scientific evaluations on the ancestry of KCB at both morphological and molecular levels.

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Species identification of the Anopheles kyrcanus complex found in Korea using PCR (PCR을 이용한 우리나라에서 발견되는 얼룩날개모기속 모기의 종 동정)

  • Yong, Tae-Sun;Lee, Han-Il;Lee, In-Yong;Lee, Jong-Won;Hwang, Ui-Uk
    • Journal of Korea Association of Health Promotion
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    • v.4 no.1
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    • pp.68-74
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    • 2006
  • For identification of four sibling species of the Anopheles hyrcanus complex found in Korea, the 5.8 rDNA-ITS2-28S rDNA region of each species was sequenced and the species-specific primers wee designed The amplified PCR products obtained from each species were analyzed by agarose gel electrophoresis. The result showed a single species- specific band, I.e. 559bp, 432bp, 322bp and 192bp for An. sinensis, An. sp., An. lesteri and An. pullus, respectively. In conclusion, the species-specific PCR primers designed from ITS2 variable regions functioned successfully and specifically, and can be applied as a useful tool for identifying species of the Anopheles hyrcanus complex found in Korea.

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Molecular Taxonomy of Ganoderma cupreum from Southern India Inferred from ITS rDNA Sequences Analysis

  • Kaliyaperumal, Malarvizhi
    • Mycobiology
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    • v.41 no.4
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    • pp.248-251
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    • 2013
  • Ganoderma is a cosmopolitan wood-rot basidiomycete that has been extensively studied for its pathogencity and medicinal properties. Identification of Ganoderma based on macro-microscopic features led to large number of synonyms which resulted in 250 taxonomic names. A Ganoderma species collected from Courtallam, Tamil Nadu was identified as G. cupreum. Phylogenetic analysis inferred from internal transcribed spacer rDNA region resolved the Indian isolate MYC1 as Ganoderma cupreum which clustered with Australian and Asian "cupreum" clade with 85% bootstrap support BS and shared 99% and 98% nucleotide similarity with Malaysian and Australian 'cupreum' respectively. This study represents the first molecular evidence of G. cupreum from Asian origin.

Degradation of Malic Acid by Issatchenkia orientalis KMBL 5774, an Acidophilic Yeast Strain Isolated from Korean Grape Wine Pomace

  • Seo, Sung-Hee;Rhee, Chang-Ho;Park, Heui-Dong
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.521-527
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    • 2007
  • Several yeast strains degrading malic acid as a sole carbon and energy source were isolated from Korean wine pomace after enrichment culture in the presence of malic acid. Among them, the strain designated as KMBL 5774 showed the highest malic acid degrading ability. It was identified as Issatchenkia orientalis based on its morphological and physiological characteristics as well as the nucleotide sequences of the internal transcribed spacer (ITS) 1-5.8S rDNA-ITS II region. Phylogenetic analysis of the ITS I-5.8S rDNA-ITS II sequences showed that the KMBL 5774 is the closest to I. orientalis zhuan 192. Identity of the sequences of the KMBL 5774 was 99.5% with those of I. orientalis zhuan 192. The optimal pH of the media for the growth and malic acid degradation by the yeast was between 2.0 and 3.0, suggesting that the strain is an acidophile. Under the optimized conditions, the yeast could degrade 95.5% of the malic acid after 24 h of incubation at $30^{\circ}C$ in YNB media containing 2% malic acid as a sole carbon and energy source.

Development of RT-PCR Kit for Diagnosis of Pathogenic Agent of Ginseng Root Rot in the Ginseng Field (인삼포장에서 뿌리섞음병원균의 진단을 위한 RT-PCR KIT의 개발)

  • 도은수
    • Korean Journal of Plant Resources
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    • v.16 no.1
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    • pp.40-48
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    • 2003
  • Cylindrocarpon destructans is the major pathogen inducing the root rot disease in ginseng. Up to now, there is no reliable and convenient method to analyze the spore density or population of this pathogen in ginseng-growing soil or any contaminated farmlands. Therefore, it will be very valuable to develop a new and reliable method in detecting the spore of this pathogen. In this study, a molecular biological technique using two step nested PCR method, was developed. Two universal ITS primers, ITS5F and ITS4R were used in the first round of PCR to amplify a fragment of ITS region from the genomic DNA of C. destructans. The specific prmers Nest 1 and Nest 2 were designed and used in the second round of PCR to amplify a inner fragment from the first round PCR product of C. destructans. C. destructans spore, only soil samples from the diseased ginseng farm produced the positive bands, suggesting its usefulness in detecting the C. destructans spores in soil samples. Thus it is recommended to first extract the whole genomic DNA from soil samples and use it for the PCR reaction, thereby eliminating the inhibitory activity of soil components.

Molecular Identification of Zoysia japonica and Zoysia sinica (Zoysia Species) Based on ITS Sequence Analyses and CAPS (ITS 염기서열 분석 및 CAPS를 이용한 조이시아 속(Zoysia) 들잔디와 갯잔디의 구별)

  • Hong, Min-Ji;Yang, Dae-Hwa;Jeong, Ok-Cheol;Kim, Yang-Ji;Park, Mi-Young;Kang, Hong-Gyu;Sun, Hyeon-Jin;Kwon, Yong-Ik;Park, Shin-Young;Yang, Paul;Song, Pill-Soon;Ko, Suk-Min;Lee, Hyo-Yeon
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.344-360
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    • 2017
  • Zoysiagrasses are important turf plants used for school playgrounds, parks, golf courses, and sports fields. The two most popular zoysiagrass species are Zoysia japonica and Zoysia sinica. These are widely distributed across different growing zones and are morphologically distinguishable from each other; however, it is phenotypically difficult to differentiate those that grow along the coastal line from those in beach area habitats. A combination of morphological and molecular approaches is desirable to efficiently identify these two plant cultivars. In this study, we used a rapid identification system based on DNA barcoding of the nrDNA-internal transcribed spacer (ITS) regions. The nrDNA-ITS regions of ITS1, 5.8S nrDNA, and ITS2 from Z. japonica, Z. sinica, Agrostis stolonifera, and Poa pratensis were DNA barcoded to classify these grasses according to their molecular identities. The nrDNA-ITS sequences of these species were found at 686 bp, 687 bp, 683 bp, and 681 bp, respectively. The size of ITS1 ranged from 248 to 249 bp, while ITS2 ranged from 270 to 274 bp. The 5.8S coding region ranged from 163 - 164bp. Between Z. japonica and Z. sinica, nineteen (2.8%) nucleotide sites were variable, and the G+C content of the ITS region ranged from 55.4 to 63.3%. Substitutions and insert/deletion (indel) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted to cleaved amplified polymorphic sequence (CAPS) markers, and applied to the Zoysia grasses sampled to verify the presence of these markers. Among the 62 control and collected grass samples, we classified three groups: 36 Z. japonica, 22 Z. sinica, and 4 Z. japonica/Z. sinica hybrids. Morphological classification revealed only two groups; Z. japonica and Z. sinica. Our results suggest that used of the nrDNA-ITS barcode region and CAPS markers can be used to distinguish between Z. japonica and Z. sinica at the species level.

Genetic Relationships of Four Korean Oysters Based on RAPD and Nuclear rDNA ITS Sequence Analyses

  • Kim, Woo-Jin;Lee, Jeong-Ho;Kim, Kyung-Kil;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee-Jeong;Jung, Hyung-Taek
    • The Korean Journal of Malacology
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    • v.25 no.1
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    • pp.41-49
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    • 2009
  • Random amplified polymorphic DNA (RAPD) marker and sequence analyses of the internal transcribed spacer (ITS) region of ribosomal DNA were used to assess phylogenetic relationships of four Korean oyster species. The average number of species-specific markers identified from five universal rice primers (URPs) by RAPD-PCR was 1.8 for Crassostrea gigas, 3.2 for C. nippona, 3.6 for C. ariakensis, and 4.6 for Ostrea denselamellosa. The length of the ITS (ITS1-5.8S-ITS2) region ranged from 1,001 to 1,206 bp (ITS1, 426-518 bp; 5.8S, 157 bp; and ITS2, 418-536 bp), while the GC content ranged from 55.5-61.1% (ITS1, 56.8-61.8%; 5.8S, 56-57.3%; and ITS2, 54.1-62.2%). A phylogenetic analysis of the oysters based on our RAPD, ITS1, and ITS2 sequence data revealed a close relationship between C. gigas and C. nippona and a distant relationship between the genera Crassostrea and Ostrea. Our results indicated that RAPD and ITS sequence analysis was a useful tool for the elucidation of phylogenetic relationships and for the selection of species-specific markers in Korean oysters.

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