• Title/Summary/Keyword: 16S and 26S rRNA genes

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Studies on the riboxomal RNA genes of rhizobium meliloti and bradyrhizobium japonicum (Rhizobium meliloti와 bradyrhizobium japonicum의 ribosomal RNA 유전자에 관한 연구)

  • 강홍규;김달웅;하지홍
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.312-317
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    • 1988
  • The genes for ribosomal RNA in Rhizobium meliloti and Bradyrhizobium japonicum were analyzed by southern hybridization of BamHI, EcoRI, HindIII digested chromosomal DNA with purified 5' $^{32}P$-labeled 16S and 23S rRNA. The big differences in the hybridization pattern of both rhizobia were found. The comparative results were discussed in relation to the copy number and conservativity of restriction sites in the rRNA genes of both rhizobia.

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Simple and Rapid Detection of Vancomycin-Resistance Gene from Enterococci by Loop-Mediated Isothermal Amplification

  • Baek, Yun Hee;Hong, Seung Bok;Shin, Kyeong Seob
    • Biomedical Science Letters
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    • v.26 no.3
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    • pp.149-156
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    • 2020
  • We developed a simple and rapid method for detecting vancomycin resistance genes, such as vanA and vanB, using loop-mediated isothermal amplification (LAMP). To identify not only vancomycin resistance genes, but also the genus Enterococcus, primers were designed for vanA, vanB, and 16S rRNA. Screening for vancomycin susceptibility in Enterococcus was performed using Etest (bioMérieux Inc). The results of the LAMP assay were compared to those of real-time RT-PCR. The optimal conditions for the LAMP assay were 65℃ for 60 min. The detection limits of the LAMP assay for vanA, and vanB were 2 × 102 copies/reaction. Compared to RT-PCR, the sensitivities and specificities of LAMP for 16S rRNA, vanA, and vanB were 100/100%, 100/100%, and 100/100%, respectively. The vanA genotype-vanB phenotype accounted for 57.5% (46/80) of the vancomycin-resistant Enterococci samples collected from 2016 to 2019. In conclusion, the LAMP assay developed in this study showed high sensitivity and specificity for vancomycin-resistant genes. Moreover, due to the simplicity and rapidity of the LAMP assay, its use can be very useful in clinical microbiology laboratories.

Characterization of Photobacterium damselae subsp. piscicida isolated from cultured starry flounder, Platichthys stellatus in Korea (우리나라 양식 강도다리, Platichthys stellatus에서 분리된 Photobacterium damselae subsp. piscicida의 특성)

  • Cho, Young Ah;Han, Hyun-Ja;Mun, Hee Eun;Jung, Sung Hee;Park, Myoung Ae;Kim, Jin Woo
    • Journal of fish pathology
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    • v.26 no.2
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    • pp.77-88
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    • 2013
  • Starry flounder, Platichthys stellatus (body length $4.4{\pm}0.51cm$) that became sick during an outbreak of disease at mariculture facilities at Ulsan, Korea in August of 2012, were examined to identify the cause of the disease. Diseased fish didn't show a unique sign, but the oxidase-positive and gram negative rod was isolated from moribund fish. The bacterium was revealed as Photobacterium damselae subsp. piscicida by biochemical analysis and sequence analysis of the 16S rRNA and capsular polysaccharide (CPS) genes. The isolates (AD5) was carrying susceptible to ofloxacin and gentamycin and showed high growth value at $18^{\circ}C$ and $25^{\circ}C$ compared to four other P. damsela strains.

Genetic identification of Aeromonas species using a housekeeping gene, rpoD, in cultured salmonid fishes in Gangwon-Do (강원도 양식 연어과 어류에서 분리된 에로모나스 종의 유전학적 동정)

  • Lim, Jongwon;Koo, Bonhyeong;Kim, Kwang Il;Jeong, Hyun Do;Hong, Suhee
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.79-88
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    • 2017
  • At the present, fish farms are suffering a lot of economic losses due to infectious diseases caused by various pathogens including aeromonad. Aeromonad is ubiquitous bacteria that causes infectious diseases. At least 26 species in the genus Aeromonas have been reported to cause fatal infections not only in salmonid fishes, but also in other freshwater and seawater fishes. Molecular techniques based on nucleic acid sequences of 16S rDNA and housekeeping genes can be used to identify the Aeromonas species. In this study, The genus Aeromonas was isolated from salmonid fishes of sixteen fish farms in Gangwon-Do, Korea and phylogenetically identified based on the sequences of 16S rDNA and housekeeping genes for Aeromonad, i.e. RNA polymerase sigma factor ${\sigma}^{70}$ (rpoD) or DNA gyrase subunit B (gyrB). Consequently, 96 strains were collected from Atlantic salmon (Salmo salar), coho salmon (Oncorhynchus kisutch), masou salmon (Oncorhynchus masou) and rainbow trout (Oncorhynchus mykiss), and 36 isolates were identified as the genus Aeromonas by 16S rDNA analysis. Thirty six Aeromonad isolates were further analysed based on rpoD or gyrB gene sequences and found Aeromonas salmonicida (24 isolates), A. sobria (10 isolates), A. media (1 isolates) and A. popoffii (1 isolates), indicating that A. salmonicida is a main infectious bacteria in Salmonid fishes in Gangwon-Do. It was also proved that the phylogenetic identification of Aeromonas species based on the sequences of housekeeping gene is more precise than the 16S rDNA sequence.

Genomic Analysis of the Extremely Halophilic Archaeon Halobacterium noricense CBA1132 Isolated from Solar Salt That Is an Essential Material for Fermented Foods

  • Lim, Seul Ki;Kim, Joon Yong;Song, Hye Seon;Kwon, Min-Sung;Lee, Jieun;Oh, Young Jun;Nam, Young-Do;Seo, Myung-Ji;Lee, Dong-Gi;Choi, Jong-Soon;Yoon, Changmann;Sohn, Eunju;Rahman, MD. Arif-Ur;Roh, Seong Woon;Choi, Hak-Jong
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1375-1382
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    • 2016
  • The extremely halophilic archaeon Halobacterium noricense is a member of the genus Halobacterium. Strain CBA1132 (= KCCM 43183, JCM 31150) was isolated from solar salt. The genome of strain CBA1132 assembled with 4 contigs, including three rRNA genes, 44 tRNA genes, and 3,208 open reading frames. Strain CBA1132 had nine putative CRISPRs and the genome contained genes encoding metal resistance determinants: copper-translocating P-type ATPase (CtpA), arsenical pump-driving ATPase (ArsA), arsenate reductase (ArsC), and arsenical resistance operon repressor (ArsR). Strain CBA1132 was related to Halobacterium noricense, with 99.2% 16S rRNA gene sequence similarity. Based on the comparative genomic analysis, strain CBA1132 has distinctly evolved; moreover, essential genes related to nitrogen metabolism were only detected in the genome of strain CBA1132 among the reported genomes in the genus Halobacterium. This genome sequence of Halobacterium noricense CBA1132 may be of use in future molecular biological studies.

Isolation of Mannanase-producing Bacteria, Bacillus subtilis WL-6 and WL-11, and Cloning and Characterization of Mannanase (Bacillus subtilis 분리균 2주 유래 mannanases의 특성 비교)

  • Yoon, Ki-Hong
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1113-1120
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    • 2016
  • Two bacterial strains producing extracellular man nanase were isolated from doenjang, a traditionally fermented soybean paste in Korea. The isolates, WL-6 and WL-11, were identified as Bacillus subtiis on the basis of their 16S rRNA gene sequences, morphological, and biochemical properties. Two genes encoding the mannanase of both B. subtilis WL-6 and B. subtilis WL-11 were each cloned into Escherichia coli, and their nucleotide sequences were determined. Both mannanase genes consisted of 1,086 nucleotides, encoding polypeptides of 362 amino acid residues. The deduced amino acid sequences of the two WL-6 and WL-11 mannanases, designated Man6 and Man11, respectively, differed from each other by eight amino acid residues, and they were highly homologous to those of mannanases belonging to the glycosyl hydrolase family 26. The 26 amino acid stretch in the N-terminus of Man6 and Man11 was a predicted signal peptide. Both Man6 and Man11 were localized at the level of 94–95% in an intracellular fraction of recombinant E. coli cells. The enzymes hydrolyzed both locust bean gum and mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, forming mannobiose and mannotriose as predominant products. The optimal reaction conditions were 55°C and pH 6.0 for Man6, and 60°C and pH 5.5 for Man11. Man11 was more stable than Man6 at high temperatures.

Sequence Divergence and Phylogenetic Investigation of the Nymphalidae (Lepidoptera: Papilionoidea) Occurring in South Korea

  • Wan, Xinlong;Kim, Min Jee;Cho, Youngho;Jun, Jumin;Jeong, Heon Cheon;Lee, Kwang Youll;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.26 no.2
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    • pp.95-112
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    • 2013
  • As a first step toward understanding the divergence and relationships of the Nymphalidae (Lepidoptera: Papilionoidea) occurring in South Korea, cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), and elongation factor-$1{\alpha}$ (EF-$1{\alpha}$) that comprise 3,501-3,716 bp were either sequenced (55 species) or the sequences were obtained from GenBank (23 species). The concatenated sequence divergence of six nymphalid subfamilies ranked in the following order: Danainae (10.3%), Satyrinae (9.5%), Limenitidinae (8.0%), Apaturinae (7.0%), Nymphalinae (6.7%), and Heliconiinae (6.2%). As has been reported in previous large scale international studies, the subfamilial relationships of (((((Limenitidinae + Heliconiinae) + (Nymphalinae + Apaturinae)) + Satyrinae) + Libytheinae) + Danainae) were also confirmed, except for the switched positions between Danainae and Libytheinae, and supported all subfamilies and tribe monophylies. Unlikely consistent phylogenetic relationships among genera within the majority of tribes in Nymphalidae, a conflicting relationship within the subfamily Apaturinae was obvious, presenting Apatura as sister to either Mimathyma or (Mimathyma + (Sephisa + (Hestina + Sasakia))), and both of these relationships are unconventional. Within the subfamily Limenitidinae, the genus Neptis was consistently revealed as a paraphyletic with respect to the genus Aldania, requiring further taxonomic investigation of the genus. Although limited, current sequence information and phylogenetic relationships are expected to be helpful for further studies.

Effects of the Antibiotics Growth Promoter Tylosin on Swine Gut Microbiota

  • Kim, Jungman;Guevarra, Robin B.;Nguyen, Son G.;Lee, Ji-Hoon;Jeong, Dong Kee;Unno, Tatsuya
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.876-882
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    • 2016
  • Tylosin has been used as a livestock feed additive and antibiotic growth promoter for many years. However, the mode of action by which tylosin enhances animal growth is unclear. We used high-throughput sequencing of 16S rRNA genes to investigate the effects of tylosin as a feed additive on swine gut microbiota. No significant difference in the rate of weight increase was observed between control and tylosin-treated pigs during a 10-week feeding trial. However, tylosin-treated pigs showed rapid increases in the relative abundance of the phylum Firmicutes. Increases in Firmicutes species are associated with (so-called) obese-type gut microbiota. The abundance of species of four families of the phylum Firmicutes (Streptococcaceae, Peptococcaceae, Peptostreptococcaceae, and Clostridiaceae) correlated positively with host weight gain. The abundance of Streptococcaceae family bacteria was least affected by tylosin treatment. Distribution analysis of operational taxonomic units (OTUs) showed that both control and tylosin-treated pigs exhibited similar OTU alterations during growth. However, the tylosin-treated group showed distinctive alterations in gut microbiota when the host weighed approximately 60 kg, whereas similar alterations occurred at around 80 kg in the control group. Our results suggest that use of tylosin accelerates maturation of swine gut microbiota rather than altering its composition.

Microbial Diversity in the Enrichment Cultures from the Fermented Beverage of Plant Extract Using Ribosomal RNA Sequence Analysis (라이보좀 RNA 염기서열 분석을 이용한 집식배양된 식물추출물발효음료의 미생물 다양성)

  • Lee, Choung Kyu;Kim, Baolo;Kang, Young Min;Lee, Hee Yul;Hwang, Chung Eun;Ahn, Min Ju;Seo, Weon Taek;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.351-359
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    • 2014
  • A beverage was produced by the fermentation of mixed extracts from the various fruits, vegetables, algae, and medical herbs. The physicochemical properties of the fermented beverage of plant extracts (FBPE) and microbial diversity were analyzed in cultures enriched from FBPE using 16S and 26S rRNA gene sequence analyses. The pH, acidity, $^{\circ}brix$, reducing sugar, and alcohol contents of the FBPE were determined to be the 3.48, 1.68%, 70.0, 1,026 g/L, and 3.5%, respectively. The most abundant free sugar and organic acid in the FBPE were glucose (567.83 g/L) and tartaric acid (93.68 mg/L), respectively. Lactobacillus homohiochii was the predominant species in all enriched culture samples: 100% of the species in 0B (0% sugar) and 40B (40% sugar) libraries and 95.6% of 20B library (20% sugar). Lactobacillus fructivorans was detected in the 20B library. The predominant species in the samples of enrichment cultures collected from FBPE with three different sugar concentrations were: Candida zeylanoides (45.2%) in the 0Y library (0% sugar), Candida lactis-condensi (35.7%) and C. zeylanoides (35.7%) in the 20Y library (20% sugar), and C. lactis-condensi (38.1%) in the 40Y library (40% sugar). This result may provide a useful frame of reference for further analyses of microbial population dynamics in FBPE.

Phylogenetic Analysis of 680 Prokaryotes by Gene Content (유전자 보유 계통수를 이용한 원핵생물 680종의 분석)

  • Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.26 no.6
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    • pp.711-720
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    • 2016
  • To determine the degree of common genes and the phylogenetic relationships among genome-sequenced 680 prokaryotes, the similarities among 4,631 clusters of orthologous groups of protein (COGs)’ presence/ absence and gene content trees were analyzed. The number of COGs was in the range of 103–2,199 (mean 1377.1) among 680 prokaryotes. Candidatus Nasuia deltocephalinicola str. NAS-ALF, an obligate symbiont with insects, showed the minimum COG, while Pseudomonas aeruginosa PAO1, an opportunistic pathogen, represented the maximum COG. The similarities between two prokaryotes were 49.30–99.78 % (mean 72.65%). Methanocaldococcus jannaschii DSM 2661 (hyperthermophilic and autotrophic, Euryarchaeota phylum) and Mesorhizobium loti MAFF303099 (mesophilic and symbiotic, alpha-Proteobacteria class) had the minimum amount of similarities. As gene content may represent the potential for an organism to adapt to each habitat, this may represent the history of prokaryotic evolution or the range of prokaryotic habitats at present on earth. COG content trees represented the following. First, two members of Chloroflexi phylum (Dehalogenimonas lykanthroporepellens BL-DC-9 and Dehalococcoides mccartyi 195) showed a greater relationship with Archaea than other Eubacteria. Second, members of the same phylum or class in the 16S rRNA gene were separated in the COG content tree. Finally, delta- and epsilon-Proteobacteria were in different lineages with other Proteobacteria classes in neighbor-joining (NJ) and maximum likelihood (ML) trees. The results of this study would be valuable to identifying the origins of organisms, functional relationships, and useful genes.