• Title/Summary/Keyword: intergenic spacer 1

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Genetic Relationship of the Ampelopsis brevipedunculata var. heterophylla and Vitis thunbergii var. sinuata with the Other Vitis Plants (개머루와 까마귀머루의 유전적 유연관계 분석)

  • Bae, Young-Min
    • Journal of Life Science
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    • v.27 no.1
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    • pp.89-94
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    • 2017
  • DNA sequences of the intergenic spacer 1 and intergenic spacer 2 of the nineteen plants belonging Vitis genus were collected from the Genbank. DNA sequences of the same regions of Vitis thunbergii var. sinuata and Ampelopsis brevipedunculata var. heterophylla, both common plants in Korea, were not available in Genbank. Those two plants were collected, their genomic DNA encoding 18S rRNA, intergenic spacer 1, 5.8S rRNA, intergenic spacer 2 and part of 28S rRNA amplified and DNA sequence determined. DNA sequences of twenty-one plants including two Korean plants were aligned by the Multiple sequence comparison by log-expectation(MUSCLE) algorithm and the alignment was used to calculate neighbor-joining tree and pairwise distance. The results indicate DNA sequences of the two Korean plants are highly homologous with each other, but they are quite distantly related to the other Vitis plants. Distant relationship of the two Korean plants with the other Vitis plants might be due to independent evolution of those two plants in geographically isolated environment. Those two Korean plants are classified in different genera based on the morphology, one in Vitis genus and the other in Ampelopsis genus, providing another example of discrepancy between morphological and genetic classification.

Use of 16S-23S rRNA Intergenic Spacer Region for identification in the fish pathogenic Streptococcus iniae (16S-23S rRNA Intergenic Spacer Region을 이용한 어류 병원성Streptococcus iniae의 분자생물학적 동정)

  • Jeong, Yong-Uk;Gang, Bong-Jo;Park, Geun-Tae;Heo, Mun-Su
    • Journal of fish pathology
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    • v.17 no.2
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    • pp.91-98
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    • 2004
  • This study was performed for the identification of Streptococcus sp. from cultured flounders (Paralichthys olivaceus) showing streptococcosis in the Jeju island. We isolated 10 strains of Streptococcus iniae from the cultured olive flounders with streptococcosis. Isolated strains were identified in S. iniae since they have formed the expected band through performing PCR assay using specific primers, Sin-1 (5'-CTAGAGTACACATGTACT(AGCT)AAG-3') and Sin-2 (5'-GGATTTTCCACTCCCATTAC-3'). In addition to 16S-23S rRNA intergenic spacers (ISR), operon structure of isolated strains showed that all strains had three 16S-23S rRNA ISR band patterns. The 16S-23S rRNA ISR sequence of isolated strains showed 96% sequence identity with S. iniae (GenBank accession number AF 048773). This paper is the first report that S. iniae is associated with streptococcosis of Olive flounder in Korea.

Secondary Structure of the Ribosomal Internal Transcribed Spacer (ITS) Region of Hypsizygus marmoreus (느티만가닥버섯의 ITS (internal transcribed spacer) 영역의 2차구조 분석)

  • Woo, Ju-Ri;Yoon, Hyeokjun;You, Young-Hyun;Lee, Chang-Yun;Kong, Won-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1260-1266
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    • 2013
  • The ribosomal DNA (rDNA) clusters of Hypsizygus marmoreus 3-10 and H. marmoreus 1-1 were analyzed in this study. The small subunit (SSU) and intergenic spacer 2 (IGS 2) was partially sequenced. The internal transcribed spacer 1 (ITS 1), 5.8S, internal transcribed spacer 2 (ITS 2), large subunit (LSU), intergenic spacer 1 (IGS 1), and 5S were completely sequenced. The rDNA clusters of H. marmoreus 3-10 and H. marmoreus 1-1 were 7,049 bp in length. The sequence of SSU rDNA, which corresponded to 18S rDNA, was 1,796 bp in length, and the sequence of LSU rDNA, which corresponded to 28S rDNA, was 3,348 bp in length. The ITS region that variable region and IGS region that non-transcribed spacer was 462 bp and 1,290 bp in length. The sequence of 5.8S rDNA and 5S rDNA was 153 bp and 43 bp in length, respectively. The 17 bp of the rDNA cluster in the H. marmoreus 3-10 strain was different to that in the H. marmoreus 1-1 strain, with 2 bp in the SSU, 3 bp in the ITS, 9 bp in the LSU, and 3 bp in the IGS. The analysis of the secondary structure revealed that the ITS regions of H. marmoreus 3-10 and H. marmoreus 1-1 have five stem-loop structures. Interestingly, among these structures, one different nucleotide sequence resulted in a different secondary structure in stem-loop V.

Cyanobacterial Diversity Analysis Using cpcBA-Intergenic Spacer Region (cpcBA-Intergenic Spacer Region을 이용한 Cyanobacteria의 다양성 분석)

  • Choi Gang-Guk;Park Yong-Ha;Ahn Chi-Yong;Bae Myoung-Sook;Oh Hee-Mock
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.287-292
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    • 2005
  • The cyanobacterial diversity was analyzed by restriction fragment length polymorphism (RFLP) of PCR-amplified rpcBA-Intergenic Spacer (IGS) genes and cpcBA-IGS gene sequencing with a sample collected at Chuso-ri in Daechung Reservoir on March 15, 2005, The Shannon-Weiner diversity index was 0.65, indicating that the cyanobacterial community structure was simple. PCR-RFLP profiles obtained were Phormidium spp. (58 clones), Anabaena spp. (14 clones), Microcystis spp. (4 clones), Spirulina sp. (1 clone) and uncultured cyanobacteria (2 clones). The PCR-RFLP of cpcBA-IGS revealed that Phormidium spp. and Anabaena spp. dominated in the invested sample. As a consequence, it seems that the analysis of functional genes such as cpcBA-IGS can be used for the species identification and community analysis of cyanobacteria.

Phylogenetic Analysis of Nuclear Ribosomal DNA Intergenic Spacer (IGS) I Region of Phellinus linteus (Nuclear Ribosomal DNA Intergenic Spacer(IGS) I 영역의 분석에 의한 목질진흙버섯의 계통분류학적 위치)

  • Rew, Young-Hyun;Lee, Jin-Hyung;Kim, Jong-Guk
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.148-151
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    • 2004
  • This study was carried out to elucidate phylogenetic relationship of a yellow lump, Phellinus linteus by comparing the nuclear ribosomal intergenic spacer (IGS) I region with that of other genera of basidiomycetes retrieved from Genbank. IGS I region of Phellinus linteus was 730 bp long and sequence homology was conserved in the 5' region, in particular $1{\sim}280\;bp$, and decreased in the direction toward the 3' end. ITS region was widely studied in phylogenies related to basidiomycetes, but IGS region was not well understood yet. Our study indicated that IGS region can be a good tool in phylogenetic study of basidiomycetes.

Detection of Pectobacterium chrysanthemi Using Specific PCR Primers Designed from the 16S-23S rRNA Intergenic Spacer Region

  • Kwon, Soon-Wo;Myung, In-Sik;Go, Seung-Joo
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.252-256
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    • 2000
  • The 16S-23S rRNA intergenic spacer regions (ISRs) were sequenced and analyzed to design specific primer for identification of Pectobacterium chrysanthemi. Two types ISRs, large and small ISRs, were identified from three strains (ATCC 11663, KACC 10163 and KACC 10165) of P. chrysanthemi and Pectobacterium carotovorum subsp. carotovorum ATCC 15713.Large ISRs contained transfer RNA-Ile(tRNA$^{Ile}$)and tRNA$^{Ala}$, and small ISRs contained tRNA$^{Glu}$. Size of the small ISRs of P. chrysanthemi ranged on 354-356 bp, while it was 451 bp in small ISR of P. carotovorum subsp. carotovorum ATCC 15713. From hypervariable region of small ISRs, species-specific primer for P. chrysanthemi with 20 bp length (CHPG) was designed from hypervariable region of small ISRs, which was used as forward promer to detect P. chrysanthemi strains with R23-1R produced PCR product of about 260bp size (CHSF) only from P. chrysanthemi strains, not from other Pectobacterium spp. and Erwinia spp. Direct PCR from bacterial cell without extracting DNA successfully amplified a specific fragment, CHSF, from P. chrysanthemi ATCC 11663. The limit of PCR detection was 1${\pm}10^2$ cfu/ml.

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Use of 16S-23S rRNA Intergenic Spacer Region for Species-specific Primer Developed of Vibrio Ichthyoenteri (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio ichthyoenteri Species-specific Primer 개발)

  • Moon Young-Gun;Heo Moon-Soo
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.117-124
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    • 2005
  • Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

Intraspecific sequence variation of trnL/F intergenic region (cpDNA) in Sedum takesimense Nakai (Crassulaceae) and aspects of geographic distribution (섬기린초에서 엽록체 DNA 염기서열의 종내 변이와 지리적 분포 양상 연구)

  • Lee, Woong;Pak, Jae-Hong
    • Korean Journal of Plant Taxonomy
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    • v.40 no.3
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    • pp.157-162
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    • 2010
  • Sequences of the trnL/F intergenic spacer of chloroplast DNA were used to investigate the intraspecific evoution and phylogeography of Sedum takesimense (Crassulaceae). The trnL/F intergeneric spacer sequences from 32 individuals of S. takesimense were either 291 bp (17 samples "without indel" in the following) or 297 bp (15samples "with indel 1") in length due to an indel of 6 bp. Two main cpDNA haplotypes were detected within S. takesimense. The haplotype with indel was found on Ulleung Island and without indel on Ulleung Island and Dok Island. This confirmed the existence of two cpDNA lineages with different geographical distributions. The cpDNA sequence analysis also suggested a putative long-distance dispersal event between Ulleung Island and Dok Island.

Cloning and Characerization of the Ribosomal RNA Gene from Gonyaulax polyedra

  • Lee, Hee-Gyun;Lee, Ji-Yeon;Lee, Dong-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.515-523
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    • 2001
  • The dinoflagellates have some primitive nuclear features and are evolutionarily intermediate between prokaryotes and eukaryotes. The small subunit ribosomal RAN gene, the 5.8S ribosomal RNA gene, and the internal transcribed spacer (ITS) of Gonyaulax polyedra were cloned, and their sequences were analyzed to better understand their evolutionary position. The small subunit ribosomal RNA gene was 1,794 nt long, the large subunit ribosomal RNA gene was approximately 3,500 nt long, and the 5.8S ribosomal RNA gene was 159 nt long. The first internal transcribed spacer (ITS1) was 191 nt long, and the second internal transcribed spacer (ITS2) was 185 nt long. The intergenic spacer of the ribosomal RNA gene (IGS) was about 2,200 nt long, indicating that 5,800 nt of transcribed sequences were separated by roughly 2,200 nt of intergenic spacer. The ribosomal RNA genes were repeated many times and arranged in a head-to-tail, tandemly repeated manner. The repeating unit of ribosomal RNA gene of G. polyedra was proposed to be 8,000 nt long. Based on the lengths of ribosomal RNA, sequence alignments with representative organisms, and phylogenetic analysis on ribosomal RNA, G. polyedra appears to be one of the alveolates branched from the eukaryotic crown and, among dinoflagellates, it seems to not have emerged early.

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PCR-RFLP Analysis of Ribosomal DNA Intergenic Spacer Region in Fusarium section Liseola. (Fusarium section Liseola 균주들에서 rDNA Intergenic Spacer 부위의 PCR-RFLP 분석)

  • 이경은;최영길;민병례
    • Korean Journal of Microbiology
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    • v.38 no.1
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    • pp.7-12
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    • 2002
  • The intergenic spacer (IGS) region of the ribosomal DNA of species in Fusarium section Liseola was analyzed by amplification and subsequent digestion with several restriction enzymes. The length of the amplified IGS region was about 2.6 Kb in all strains except F.moniliforme 12 Which was about 2.9 Kb. The enzymes, EcoRI, HincII, SalI, HindIII, PstI and SmaI, digested the IGS region and nine haplotypes were identified among 11 strains. In the dendrogram based on PCR-RFLP of IGS region combined the results of section Liseola in this study and section Elegans in previous study, variation in the IGS appears to offer considerable potential to resolve intraspecific relationship as well as interspecies or intersection.