• The Plant Pathology Journal
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• 제28권3호
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• pp.295-298
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• 2012

Genetic diversity among Pseudomonas syringae pv. morsprunorum isolates from Prunus mume in Korea and Japan was investigated by comparative sequence analysis of the 16S rRNA gene. The strains included 24 field isolates recovered from P. mume in Korea along with seven Japanese strains. Two strains isolated from P. salicina in Japan, one strain from P. avium in the United Kingdom, and the pathotype strain were also used for comparison with their 16S rRNA gene sequences. Nearly complete 16S rRNA gene sequences were sequenced in all 35 strains, and three sequence types, designated types I, II and III, were identified. Eleven strains consisting of five Korean isolates, five Japanese strains, and one strain from the United Kingdom belonged to type I, whereas the pathotype strain and another 19 Korean isolates belonged to type III. Another four Japanese strains belonged to type II. Type I showed 98.9% sequence homology with type III. Type I and II had only two heterogeneous bases. The 16S rRNA sequence types were correlated with the races of P. syringae pv. morsprunorum. Type I and II strains belonged to race 1, whereas type III isolates were included in race 2. Sequence analyses of the 16S rRNA gene from P. syringae pv. morsprunorum were useful in identifying the races and can further be used for epidemiological surveillance of this pathogen.

• 생태와환경
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• 제46권2호
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• pp.301-309
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• 2013

2012년 북한강수계 전역에 걸쳐 대발생을 일으킨 남조 Anabaena의 분자생물학적 특성을 검토하였다. 시료는 2012년 7월 13일에 남조 Anabaena 밀도가 높았던 3개 호소- 의암호, 청평호, 팔당호에서 각각 채집하였다. 분석은 선행연구 문헌을 근거로 하는 형태학적 분석, 16S rRNA 염기서열 분석을 통한 분자계통학적 분석을 동시에 실시하였다. 결과를 종합하면 북한강수계 3개 호소에서 조류 대발생을 일으킨 남조 Anabaena는 모두 동일종 Anabaena crassa (Lemmermann) Komark.-Legn. & Cronberg이며, 동시에 출현하고 본 종과 형태 및 분자생물학적으로 매우 유사한 A. circinalis는 환경요인에 따라 형태변이가 쉽게 일으키는 동일종으로 밝혀졌다.

• 한국임상수의학회지
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• 제31권1호
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• pp.36-39
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• 2014

Many bacteria colonized in the horse semen affect quality of the sperm and some may cause infection in the mare reproductive tract and infertility of susceptible mare. This study was initiated to determine the prevalence of bacteria in testes of Jeju horses by determining rRNA sequence. The samples were swabed from the testes of nine Jeju horses (aged from 8 to 12 months after birth). Bacteria isolated from testes were identified by 16S rDNA sequencing. 1.6-kbp PCR products for 16S rRNA coding region were obtained using the universal primers. The PCR products were further purified and sequenced. Maximum similar species were found by BLAST search in the GenBank DNA database. BLAST results showed that the sequences were similar to those of Acinetobacter sp (A. schindleri, A. ursingii)., Bacillus cereus, Corynebacterium glutamicum, Escherichia coli, Gamma proteobacterium, Micrococcus luteus, Pseudomonas mendocina, Shigella sonnei, Sphingomonas sp., Staphylococcus sp (S. cohnii, S. saprophyticus, S. xylosus)., and Stenotrophomonas maltophilia. DNA sequences for 16S rRNA is provided useful informations for species identification of pathogenic microorganisms for the reproductive organs in horses.

• 한국미생물·생명공학회지
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• 제38권3호
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• pp.263-272
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• 2010

독도 연안에 존재하는 배양 가능한 미생물의 다양성을 16S rRNA 분석으로 조사하였다. 동도 선착장 주변과 서도 숙소 부근을 중심으로 채취한 시료에서 163개의 해양 미생물을 분리하였다. 분리한 미생물 163종을 16S rRNA 염기서열의 분석을 이용하여 부분동정 할 수 있었다. 부분동정된 미생물은 gamma-proteobacteria(58%), alpha-proteobacteria (20%), bacteriodetes(16%) 계통이 대부분을 차지하고 있었고, 그 외에도 low G+C Gram positive bacteria와 epsilonproteobacteria가소수 동정 되었다. 염기서열이 분석된 미생물들은 이전에 보고된 미생물들의 16S rRNA 유전자와 93.3%에서 100%의 유사도를 보이며 56속 94종으로 부분 동정되었다. 163종의 부분 동정된 미생물 중 36개의 분리 미생물이 새로운 종으로 분류될 후보군으로 추정되었다. 본 연구의 결과 독도연안 바닷물에는 proteobacteria와 bacteriodetes의 비율이 높게 나타났고, 미생물 다양성을 높게 유지하고 있었다. 이 다양한 미생물로부터 다양한 유용미생물 자원을 확보할 수 있고, 새로운 종으로 분류될 후보군 들은 추후 여러 생리생화학적 실험을 수행하여 새로운 종 또는 새로운 속으로 발표할 수 있을 것으로 판단된다.

• 미생물학회지
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• 제41권2호
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• pp.117-124
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• 2005

Rotifer와 병든 넙치 자어로부터 분리된 2개의 균주는 표현형적인 특성 확인 결과 Vibrio ichthyoenteri로 확인이 되었다. V. ichthyoenteri를 검출하기 위래 고감도 PCR 방법 개발을 하기 위래 V. ichthyoenteri 16S-23S rRNA intergenic spacer region(ISR)을 분석하였고, V. ichthyoenteri중 특이적 primer를 개발하였다. V. ichthyoenteri 의 ISR를 분석한 결과 1개의 다형성 ISR type서열을 포함하고 있었다. ISR서열은 길이는 348bp이며 tRNA gene을 가지고 있지 않았다. 이 서열을 가지고 이미 알려진 다른 Vibrio 종의 ISR 서열과 mutiple alignment를 수행한 결과 여러 영역에서 높은 가변성을 나타내어 가변 부위를 표적으로 하여 V. ichthyoenteri를 검출하기 위한 종 특이적 primer를 제작하였다. 제작된 primer의 특이성을 확인하기 위해 Vibrio 표준균주 19종의 genomic DNA와 분리균주 18 group에 genomic DNA 그리고 V. ichthyoenteri와 가장 유사한 서열을 가지고 있다고 알려진 Vibrio 종의 genomic DNA를 가지고 시험하였다. 그 결과 본 연구에서 제작된 종 특이적 primer를가지고 PCR 반응을 하면 V. ichthyoenteri를 검출 할 수가 있다.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.504-507
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• 2001

The partial 16S gene sequences of six filamentous cyanobacterial strains, Oscillatoria lmosa KCTC AG10168, Oscillatoria princeps KCTC AG10153, Oscillatoria sp. KCTC AG 10184, Phormidium tenue KCTC AG10158, Phormidium parchydematicum KCTC AG10164, and Lyngbya hieronymusii KCTC AG10199, which were isolated in the late summer at Daecheong Reservoirs, were determined and assigned their phylogenetic and taxonomic position among taxa of order Ocillatoriales whose partial 16S rRNA gene sequences aligned in this suty, were very heterogeneously clustered with other taxa. The two strains, Oscillatoria limosa KCTC AG10168 and O. princeps KCTC AG10153, formed a cluster with O. sancta PCC7515, which supported 64% of the bootstrap trees with high similarity (19-96.15%). Strain Oscillatoria sp. KCTC AG10184, that was known to produce a nasty substance, was closely related to the toxic Oscillatoria group. The study on morphological variation in various environments and toxin production will confirm the taxonomic status of these species. Phormidium tenus KCTC AG10158 and Phormidium parchydematicum KCTC AG10164 made a cluster with other oscillatorian species of Phormidium, Oscillatoria, and Leptolynbya, which supproted 100% of the bootstrap trees with a very high sequence smilarity (96.8-99.8%) in thsi study. The sequence analysis in this study also supported that taxa of oscillatoriales are not monophyletic. Some of the fractures, such as the presence or absence of sheath and cell shape, which were used to define them, would be inadequate and should be reconfirmed. We suggest that sequences of partial 16S rRNA gene fragments aligned in this study should be more useful than morphological features in the identification and reconfirmation of the taxonomic status of these oscillactorian cyanobacteria.

• 한국미생물·생명공학회지
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• 제47권3호
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• pp.401-411
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• 2019

All Idiomarina species are isolated from saline environments; microorganisms in such extreme habitats develop metabolic adaptations and can produce compounds such as proteases with an industrial potential. ARDRA and 16S rRNA gene sequencing are established methods for performing phylogenetic analysis and taxonomic identification. However, 16S-23S ITS is more variable than the 16S rRNA gene within a genus, and is therefore, used as a marker to achieve a more precise identification. In this study, ten protease producing Idiomarina strains isolated from the Peruvian salterns were characterized using biochemical and molecular methods to determine their bacterial diversity and industrial potential. In addition, comparison between the length and nucleotide sequences of a 16S-23S ITS region allowed us to assess the inter and intraspecies variability. Based on the 16S rRNA gene, two species of Idiomarina were identified (I. zobellii and I. fontislapidosi). However, biochemical tests revealed that there were differences between the strains of the same species. Moreover, it was found that the ITS contains two tRNA genes, $tRNA^{Ile(GAT)}$ and $tRNA^{Ala(TGC)}$, which are separated by an ISR of a variable size between strains of I. zobellii. In one strain of I. zobellii (PM21), we found nonconserved nucleotides that were previously not reported in the $tRNA^{Ala}$ gene sequences of Idiomarina spp. Thus, based on the biochemical and molecular characteristics, we can conclude that protease producing Idiomarina strains have industrial potential; only two I. zobellii strains (PM48 and PM72) exhibited the same properties. The differences between the other strains could be explained by the presence of subspecies.

• Animal cells and systems
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• 제4권1호
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• pp.77-85
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• 2000

This study has demonstrated the molecular variation of 5S rRNA genes in 15 Allium subgenus Rhizirideum and 1 Allium subg. Allium. For cloning of the 5S rRNA genes, PCR products were obtained from amplification with oligonucleotide primers which were derived from the conserved coding region of 5S rRNA genes. These amplified PCR products were cloned and identified by FISH and sequence analysis. The 5S rRNA loci were primarily located on chromosomes 5 and/or 7 in diploid species and various chromosomes in alloploid species. The size of the coding region of 5S rRNA genes was 120 bp in all the species and the sequences were highly conserved within Allium species. The sizes of nontranscribed spacer (NTS) region were varied from 194 bp (A. dektiude-fustykisum, 2n=16) to 483 bp (A. sativum). Two kinds of NTS regions were observed in A. victorialis var. platyphyllum a diploid, A. wakegi an amphihaploid, A. sacculiferum, A. grayi, A. deltoide-fistulosum and A. wenescens all allotetraploids, while most diploid species showed only one NTS region. The species containing two components of NTS region were grouped with different diploid species in a phylogenetic tree analysis using the sequences of 5S rRNA genes and adjacent non-coding regions.

• 미생물학회지
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• 제47권3호
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• pp.268-274
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• 2011

남조세균 Anabaena (Cyanobacteria, Nostocales)는 담수 생태계에서 녹조 현상을 유발하거나 일부 종은 간독소(hepatotoxin)를 갖고 있어 수질관리 차원에서 주목 받아 왔다. 본 연구는 Anabaena RNA polymerase beta subunit (rpoB) 유전자 염기서열을 규명하였으며, 분류학적 분자 마커로 사용하기 위하여 이들 염기서열의 특성을 평가하였다. Anabaena rpoB 유전자는 16S rRNA 유전자와 비교하여 염기 유사도가 낮으며 유전자 변이가 큰 것으로 분석되었으며, 통계적으로 유의한 차이를 보였다(Student t-test, p<0.01). Parsimony 분석을 통해 rpoB 유전자가 4.8배의 속도로 빠르게 진화하는 것으로 파악되었다. 또한 rpoB 유전자 phylogeny 분석에서 16S rRNA tree보다 높은 해상도로 Anabaena 균주를 명확하게 구분해 주었다. 본 연구 결과는 Anabaena의 종 식별, 분자계통 분류, 분자적 검출을 위해 rpoB 유전자가 매우 효과적이라는 것을 제시해 준다.

• International Journal of Oral Biology
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• 제38권1호
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• pp.29-36
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• 2013

Mitis group streptococci (MGS) were classified based on the nucleotide sequences 16S rRNA gene (16S rDNA) and comprised 13 Streptococcus species. However, 16S rDNA homogeneity among MGS was too high to discriminate between clinical strains at the species level, notably between Streptococcus mitis, Streptococcus oralis, Streptococcus pneumoniae, and Streptococcus pseudopneumoniae. The purpose of this study was to discriminate between 37 strains of MGS isolated from Korean oral cavities using phylogenetic analysis of the DNA-dependant RNA polymerase beta-subunit gene (rpoB). 16S rDNA and rpoB from clinical strains of MGS were sequenced using the dideoxy chain termination method and analyzed using MEGA version 5 software. The resulting phylogenetic data showed that the rpoB sequences could delineate clinical strains of MGS at the species level. Phylogenetic analysis of rpoB is therefore a useful approach for identifying MGS at the species level.