• Title/Summary/Keyword: 23S rDNA

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Genetic Similarity Between Jujube Witches¡?Broom and Mulberry Dwarf Phytoplasmas Transmitted by Hishimonus sellatus Uhler

  • Cha, Byeongjin;Han, Sangsub
    • The Plant Pathology Journal
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    • v.18 no.2
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    • pp.98-101
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    • 2002
  • Using phytoplasma universal primer pair Pl and P7, a fragment of about 1.8 kb nucleotide sequences of 16S rRNA gene and 16S-23S rRNA intergenic spacer region, and a portion of 23S rRNA gene of jujube witches'broom (JWB) and mulberry dwarf(MD) phytoplasmas were determined. The nucleotide sequences of JWB and MD were 1,850 bp and 1,831 bp long, respectively. The JWB phytoplasma sequence was aligned with the homologous sequence of MD phytoplasma. Twenty-eight base insertions and nine base deletions were found in the JWB phytoplasma sequence compared with that of MD phytoplasma. The similarity of the aligned sequences of JWB and MD was 84.8%. The near-complete 16S rRNA gene DNA sequences of JWB and MD were 1,529 bp and 1,530 bp in length, respectively, and revealed 89.0% homology. The 16S-23S rRNA intergenic spacer region DNA sequences were 263 bp and 243 bp in lengths respectively, while homology was only 70% and the conserved tRNA-lle gene of JWB and MD was located into the intergenic space region between 16S-23S rRNA gene. The nucleotide sequences were 77 bp long in both JWB and MD, and showed 97.4% sequence homology. Based on the phylogenetic analysis of the two phytoplasmas, the JWB phytoplasma belongs to the Elm yellow phytoplasma group (16S rV), whereas, the MD phytoplasma belongs to the Aster yellow group (16S rI).

In vitro Selection of RNA Aptamers which Bind to Escherichia coli tRNAVal (대장균 tRNAVal에 결합하는 RNA Aptamer들의 시험관내 선별)

  • Jo, Bong Rae
    • Journal of the Korean Chemical Society
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    • v.46 no.2
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    • pp.157-163
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    • 2002
  • To identify RNA motifs interacting with $tRNA^{Val}$, a SELEX(Systematic Evolution of Ligands by Exponential Enrichment) was applied. Random DNA library which contains a region of ran-domized 48-mer oligonucleotide flanked by conserved sequ ence primers was transcribed into RNA pool using T7 RNA polymerase and RNA aptamers were selected with $tRNA^{Val}$ -immobilized affinity column through 14 rounds of SELEX. Some of the resulting aptamers contained a consensus sequence similar to the sequence in the loop regions of three rRNAs; C43GAAC47 sequence of 5S rRNA, G1491AAGU1495, G1379UUCC1383 sequence of 16S rRNA and C1064UUAG1068, G2110UGUA2114, C2480GACGG2485, A2600CAGU2604 sequence of 23S rRNA. These results suggest that $tRNA^{Val}$ can interact with 5S rRNA, 16S rRNA and 23S rRNA with variety in ribosome.

ITS Primers with Enhanced Specificity to Detect the Ectomycorrhizal Fungi in the Roots of Wood Plants

  • Kim, Dong-Hun;Chung, Hung-Chae;Ohga, Shoji;Lee, Sang-Sun
    • Mycobiology
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    • v.31 no.1
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    • pp.23-31
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    • 2003
  • With universal primer ITS1-F, the specific DHJ2 primer was developed to detect the Ectomycorrhizal(ECM) root tips in soil and to identify the species of ECM fungi, as based on DNA sequences of rDNA stored in GeneBank of NCBI. This primer was designed with the common sites of rDNA of Amanita and Boletus, and was also designed with several DNA programs provided by NCBI. The DNA fragments synthesized by PCR were calculated to be 1,000 to 1,200 bps of DNA located to 18s to 28s rDNA to contain two variable sites of ITS, indicating much diversities for specific species or ecotypes of ECM fungi. The primer DHJ2 reacted with the genomic DNA's extracted from the tissues of basidiocarp at the rate of 73 of 80 fungi collected produced single bands with a 1,100 bps length. The DNA fragment synthesized with the genomic DNA that extracted from eight ECM tips of Pinus densiflora was confirmed and analysized to the rDNAs of ECM in full sequences, and informed to be a ECM fungal species in the forest.

Identification by 16S rDNA Partial Sequencing of Thermophilic Bacteria with Fermentation of Pig Manure (16S rDNA 증폭에 의한 부분염기서열을 이용한 분뇨 발효 관련 고온 호기성 박테리아의 동정)

  • Kim, Myung Kil;Choi, Don Ha;Choi, In Gyu;Kim, Byung Gyu;Song, Jae Gyung
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.1
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    • pp.68-78
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    • 2006
  • The purpose of this study was to estimate the identification of thermophilic hacterial with fermentation of pig manure. To identify the characters of thermophilic bacteria related to fermentation at a high temperature condition, we selected 28 different kinds of original settling thermophilic bacteria that were sampled at different 23 areas. They were distributed 1$1{\times}10^5{\sim}10^8CFU$ at medium and the enzyme activity at $55^{\circ}C$ incubation condition, especially cellulase and a-amylase, were higher than those of $30^{\circ}C$. Partial sequencing data for 165 rDNA region were obtained from 28 samples representing 15 different genera. Bacillus subcilis, one of those bacteria, has endodermic spores at high fermented condition.

Study on the Genetic Variation of the Mitochondrial DNA in the Beet Armyworm, Spodoptera exigua (H bner), Using PCR-RFLP (PCR-RFLP를 이용한 파방나방 (Spodoptera exigua(H bner)) 미토콘트리아 DNA의 유전변이 연구)

  • 김용균;이명렬;정충렬
    • Korean journal of applied entomology
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    • v.37 no.1
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    • pp.23-30
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    • 1998
  • Restriction fragment length polymorphism (RFLP) of a DNA has been a useful tool for analyzing genetic variation. This research was performed to establish an RFLP analytic method on the mitochondrial DNA (mtDNA) of the beet armyworm, Spodoptera exigua (Hiibner). To do this, total size of the mtDNA was measured and polymerase chain reaction (PCR) primers were selected. Its mitochondrial genome size was ca. 16kb. From a serial PCR test of 29 primers refered to the compilation of Simon et al. (1994), 22 primers were selected to amplify its mtDNA fragments. These primers resulted in short (300-700 bp) or long (1000-2000 bp) DNA products which represented a total or partial sequence of each of CO-I, CO-11, Cyt-B, ND-1, 12s rRNA, 16s rRNA, and some tRNAs. PCR-RFLP was performed in some variable mtDNA regions with 8 kinds of 4bp recognizing restriction enzymes. Different populations from Andong, Kyungsan, and Sunchun did not show any restriction site polymorphisms but had some length variation in certain regions of mtDNA.

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Genetic Stock Identification of Spotted Flounder, Verasper variegatus from Yeocheun, Korea (범가자미에 대한 유전학적 동정)

  • KIM Kyung Kil;KIM Yoon;NAM Yoon Kwan;KIM Dong Soo
    • Journal of Aquaculture
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    • v.6 no.3
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    • pp.221-233
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    • 1993
  • Cell size, DNA content, chromosome and PCR-based mitochondrial 12S rRNA gene analyses were conducted to obtain basic informations for genetic stock identification of spotted flounder (Verasper variegatus) from Yeocheun, Korea. The mean erythrocytic and nuclear volumes of spotted flounder were $211.10{\mu}m^3$ and $23.03{\mu}m^3$, respectively. The haploid DNA content of this species was 0.79 pg/cell which correspond to $46.5\%$ of carp and to $22.6\%$ of mammals. Spotted flounder had the 2n = 46 acrocentric chromosomes but no heteromorphic sex chromosomes was found. Mitochondrial DNA gene for 12S ribosomal RNA was amplified by polymerase chain reaction (PCR) and the PCR products were subjected to digestion with 15 restriction endonucleases. Restriction enzyme analyses revealed that Ava I, Mae II, Sma I and Xba I had one restriction site in the mitochondrial 12S rRNA gene segment of spotted flounder, while Mae I had two. Segments of 12S rRNA gene from mitochondria in spotted flounder were sequenced and compared with channel catfish and human as controls. The 12S rRNA gene of this species was more similar to that of channel catfish than to human's.

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Isolation of Dextran-producing Leuconostoc Zactis from Kimchi

  • Kim, Bong-Joon;Min, Bong-Hee;Kim, Jeongho;Han, Hong-Ui
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.11-16
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    • 2001
  • Tentative identification of Leuconostoc lactis IH23 isolated from kimchi (a fermented vegetable product) has been described previously with 16S rDNA sequencing (Choi, 1., M. Sc. Thesis Inha Univ.1999). This strain produced the slime identified as dextran based on IR, $\^$13/C- and $^1$H-NMR spectroscopic results. Further study proved that the isolate IH23 belongs to a homogeneous genetic group with L. lactis DSM 20202$\^$T/ and L. argentinum DSM 8581$\^$T/. The results showed DNA-DNA homology of 99-100%, 16S rDNA gene sequence similarity (97.7% ), and a phylogenetic relationship although L. argentinum DSM 8581$\^$T/ had lower homology (80-91%). These data indicate that L. argentinum DSM 8581$\^$T/ and the isolate IH23 belong to an identical species with L. lactis DSM 20202$\^$T/at the genetic level, although in carbohydrate fermentation, the isolate IH23 was mast closely related to L. argentinum DSM 8581$\^$T/ and quite different from L. lactis DSM 20202$\^$T/. Here we first report the isolation of consistent phenotypic variation in Leuconostoc lactis. We also emphasize that the nomenclature of subspecies needs to be differentiated into the three strains mentioned above in Leuconostoc lactis.

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Morphology and Sequence Analysis of Nuclear 18S rDNA from the Summer Strain of Porphyra suborbiculata (Rhodophyta) in Korea (여름철 서식 한국산 홍조류 둥근돌김 (Porphyra suborbiculata)의 형태 및 18S rDNA 염기서열 분석)

  • JIN Long-Guo;KIM Myung-Sook;CHOI Jae-Suk;CHO Ji-Young;JIN Hyung-Joo;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.489-495
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    • 2000
  • The 185 ribosomal RNA gene (185 rDNA) of the marine alga Porphyra sp. 723 (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analysed. The Porphyra species was a summer strain collected on rocks in upper intertidal zone at Ikidae, Pusan on 23rd July 1999. The fronds were $1{\~}5 cm$ long, monostromatic, and orbicular or ovate shaped, They had spinulate processes at margin of the frond, Comparison of this 185 rDNA sequence with the other Forphyra species indicates that Porphyra sp. 723 has the same 185 rDNA sequence derived from Porphyra suborbiculata (NCBI access number; AB 013180) except one base pair substitution in 2327 base pairs.

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Identification and Comparison of the Nucleotide Sequence of 16S-23S rRNA Gene Intergenic Small SR(Spacer Region) of Lactobacillus rhamnosus ATCC 53103 with Those of L. casei, L. acidophilus and L. helveticus

  • Byun, J.R.;Yoon, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1816-1821
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    • 2003
  • Reliable PCR based identification of lactobacilli has been described utilizing the sequence of 16S-23S rRNA intergenic spacer region. Those sequence comparisons showed a high degree of difference in homology among the strains of L. rhamnosus, L. casei, L. acidophilus and L. helveticus whose 16S-23S rRNA intergenic small SR's sizes were 222 bp, 222 bp, 206 bp and 216 bp respectively. The sequence of 16S-23S rRNA intergenic spacer region of L. rhamnosus ATCC 53103 revealed the close relatedness to those of L. casei strains by the homology ranges from 95.4% to 97.2%. 16S-23S rRNA intergenic spacer region nucleotide sequence of L. acidophilus showed some distant relatedness with L. rhamnosus ATCC 53103 with the homology ranges from 40.3% to 41.8% and that with L. helveticus was shown to be 30% of homology, which exists at the most distant phylogenetic relatedness. The identification of species and strain of lactobacilli was possible on the basis of these results. The common sequences among the 17 strains were CTAAGGAA located in the initiating position of the DNA and some discrepancies were found between the same strains based on these results.

Molecular characterization of bacterial endosymbionts of Acanthamoeba isolates from infected corneas of Korean patients

  • Xuan, Ying-Hua;Yu, Hak-Sun;Jeong, Hae-Jin;Seol, Sung-Yong;Chung, Dong-Il;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • v.45 no.1 s.141
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    • pp.1-9
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    • 2007
  • The endosymbionts of 4 strains of Acanthamoeba(KA/E9, KA/E21, KA/E22, and KA/E23) isolated from the infected corneas of Korean patients were characterized via orcein stain, transmission electron microscopic examination, and 16S rDNA sequence analysis. Double membrane-bound, rod-shaped endosymbionts were distributed randomly throughout both the trophozoites and cysts of each of Acanthamoeba isolates. The endosymbionts of KA/E9, KA/E22, and KA/E23 were surrounded by electron-translucent areas. No lacunae-like structures were observed in the endosymbionts of KA/E21, the bacterial cell walls of which were studded with host ribosomes. Comparative analyses of the 16S rDNA sequences showed that the endosymbionts of KA/E9, KA/E22 and KA/E23 were closely related to Caedibacter caryophilus, whereas the KA/E21 endosymbiont was assigned to the Cytophaga-Flavobacterium-Bacteroides(CFB) phylum. In the 4 strains of Acanthamoeba, the hosts of the endosymbionts were identified as belonging to the Acanthamoeba castellanii complex, which corresponds to the T4 genotype. Acanthamoeba KA/E21 evidenced characteristics almost identical to those of KA/E6, with the exception of the existence of endosymbionts. The discovery of these endosymbionts from Acanthamoeba may prove essential to future studies focusing on interactions between the endosymbionts and the amoebic hosts.