• Title/Summary/Keyword: 16s rDNA

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Molecular Systematics of Tephritidae (Insecta : Diptera): Testing Phylogenetic Position of Korean Acidiella spp. (Trypetini) Using Mitochondrial 16S rDNA Sequences

  • Han, Ho-Yeon;Ro, Kyung-Eui
    • Animal cells and systems
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    • v.6 no.1
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    • pp.13-18
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    • 2002
  • Phylogenetic relationships of Korean Acidiella species were tested using mitochondrial 16S ribosomal RNA gene sequences. We used 16 published sequences as outgroup, and 10 new sequences for nine Korean Acidiella species as ingroup. The number of aligned sites was 1,281 bp, but 1,135 bp were used for the analysis after excluding sites with missing data or gaps. Among these 1,135 sites, 464 sites were variable and 340 were informative for parsimony analysis. Phylogenetic information was extracted from this data set using neighbor-joining, maximum likelihood and maximum parsimony methods and compared to a morphology-based phylogenetic hypothesis. Our molecular data suggest that: (1) the tribe Trypetini appears to be monophyletic even when the nine additional Acidiella species are added to our previous phylogenetic analysis; (2) all the Korean Acidiella species belong to the Trypeta group, but the genus Acidiella is not supported as monophyletic; (3) the close relationship of A. circumvaga, A. issikii, and A. sapporensis is supported; (4) the close relationship of A. pachypogon and two additional new Acidiella species is strongly supported; and (5) the possible presence of two or more cryptic species among the specimens previously identified as A. obscuripennis is suggested. Sequence data from the mitochondrial 16S rDNA allowed us to better understand the systematic status of Korean Acidiella species. They indicated that the current concept about the genus Acidiella is insufficient and needs to be refined further. This study also showed a few interesting relationships, that had not been recognized by morphological study alone. Based on this study, we were able to plan further experiments to analyze relationships within the Trypeta Group.

Rapid Detection of the pathogenic agent of Bacterial white enteritis of Larval and Juvenile Stages in Olive flounder (Paralichthys olivaceus) (넙치(Paralichthys olivaceus)자치어 장관백탁증(Bacterial white enteritis) 원인균의 신속 검출)

  • Mun, Yeong-Geon;Park, Geun-Tae;Son, Hong-Ju;Lee, Sang-Hyeon;Lee, Jeong-Min;Heo, Mun-Su
    • Journal of fish pathology
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    • v.17 no.3
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    • pp.159-169
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    • 2004
  • Bacterial wihte enteritis ocurred by infection of V. ichthyoenteri is a devastating disease in olive flounder (Paralichthys olivaceus) hatcheries in Korea. Since white enteritis has been a problem in aquqtic industries, necessity of a rapid detection method is increased. In an attempt to develop rapid PCR method 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. The intergenic spacers were amplified by primers complementary to conserved region of 16S and 23S rRNA genes. The intergenic spacer region between the 16S and 23S rRNA genes of V. ichthoenteri were investigated by PCR fragment length typing and DNA sequencing. Analysis of the ISR sequences showed that V. ichthyoenteri contains one types of polymorphic ISRs. The size of ISRs ranged 348bp length and not contains tRNA genes. Mutiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of Vibrio ichthyoenteri. PCR. The specific of the primer was examined using genomic DNA prepared from 19 different Vibrio species, isolated 18group Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

A newly developed consensus polymerase chain reaction to detect Mycoplasma species using 16S ribosomal RNA gene

  • Hong, Sunhwa;Park, Sang-Ho;Chung, Yung-Ho;Kim, Okjin
    • Korean Journal of Veterinary Service
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    • v.35 no.4
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    • pp.289-294
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    • 2012
  • Mycoplasmas are highly fastidious bacteria, difficult to culture and slow growing. Infections with Mycoplasma species can cause a variety of problems in living organisms and in vitro cell cultures. In this study, we investigated the usefulness of a genus-specific consensus PCR analysis method to detect Mycoplasma species. The developed consensus primer pairs MycoF and MycoR were designed specifically to amplify the 16S ribosomal RNA gene (rRNA) of Mycoplasma species by the optimized PCR system. The developed consensus PCR system effectively amplified 215 bp of Mycoplasma genus-specific region of 16S rRNA. In conclusion, we recommend this consensus PCR for monitoring Mycoplasma species in animals, human and cell culture system.

Characteristics and Diagnostic Methods of Streptococcosis Causing Disease in Aquaculture (양식 어류에 질병을 유발하는 연쇄구균증의 특성 및 진단 방법)

  • Kim, Dong-Hwi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1118-1126
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    • 2018
  • In this study, investigated the general characteristics and diagnostic methods types of streptococcosis among various fish disease pathogens that caused a lot of economic damaged to aquaculture fish based on the previous research paper. Streptococcosis infection of fish is considered a reemerging disease affecting a variety of wild and cultured fish throughout the world. Calssifiacation of Gram positive cocci based on DNA-DNA hybridization coupled with 16S sequencing has shown that at least five different species are considered of significance as fish pathogens: Lactococcus garvieae, L. piscium, Streptococcus iniae, S. agalactiae, S. paruberis, Vagococcus salmoninarum. Symptoms of infection with streptococcosis disease such as body color change, eyeball abnormality, gill discoloration, bleeding, abdominal distension, swelling of the kidney and spleen. In addition, it usually occurs from June to October when the water temperature rise a lot of fish death. Currently, 16S rRNA, 16S-23S rRNA intergenic spacer region (ISR), Random Amplified polymorphic DNA (RAPD), Ribotyion (RT), Loop-mediated isothermal amplification (LAMP) are among the methods for diagnosing streptococcosis. Among them, the LAMP method, which is high applicable to the aquaculture farm has attracted the spotlight, but due to problems such as confirmation of results. This seems to minimize the economic loss of streptococcosis which complements the problem so that it can be easily used from the diagnosis to the results confirmation.

Null Allele in the D18S51 Locus Responsible for False Homozygosities and Discrepancies in Forensic STR Analysis

  • Eom, Yong-Bin
    • Biomedical Science Letters
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    • v.17 no.2
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    • pp.151-155
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    • 2011
  • Short tandem repeats (STRs) loci are the genetic markers used for forensic human identity test. With multiplex polymerase chain reaction (PCR) assays, STRs are examined and measured PCR product length relative to sequenced allelic ladders. In the repeat region and the flanking region of the commonly-used STR may have DNA sequence variation. A mismatch due to sequence variation in the DNA template may cause allele drop-out (i.e., a "null" or "silent" allele) when it falls within PCR primer binding sites. The STR markers were co-amplified in a single reaction by using commercial PowerPlex$^{(R)}$ 16 system and AmpFlSTR$^{(R)}$ Identifiler$^{(R)}$ PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI PRISM$^{(R)}$ 3100 Genetic Analyzer with capillary electrophoresis. The GeneMapper$^{TM}$ ID software were used for size calling and analysis of STR profiles. Here, this study described a forensic human identity test in which allelic drop-out occurred in the STR system D18S51. During the course of human identity test, two samples with a homozygous (16, 16 and 21, 21) genotype at D18S51 locus were discovered using the PowerPlex$^{(R)}$ 16 system. The loss of alleles was confirmed when the samples were amplified using AmpFlSTR$^{(R)}$ Identifiler$^{(R)}$ PCR amplification kit and resulted in a heterozygous (16, 20 and 20, 21) genotype at this locus each other. This discrepancy results suggest that appropriate measures should be taken for database comparisons and that allele should be further investigated by sequence analysis and be reported to the forensic community.

Analysis of RNA Polymerase Beta Subunit (rpoB) Gene Sequences for the Discrimination of Cyanobacteria Anabaena Species (남조세균 Anabaena 종 구분을 위한 RNA Polymerase Beta Subunit (rpoB) 유전자 염기서열 분석)

  • Cheon, Ju-Yong;Lee, Min-Ah;Ki, Jang-Seu
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.268-274
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    • 2011
  • Anabaena (Cyanobacteria, Nostocales) are important for water quality controls, because they are often responsible for freshwater green tides; moreover, some species are reported to produce hepatotoxin. In this study, we sequenced RNA polymerase beta subunit (rpoB) gene of Anabaena, and evaluated their sequences for the potential use of a molecular taxonomic marker in this taxon. Anabaena rpoB showed low DNA similarity and high genetic divergences when compared those of 16S rRNA, and the molecular differences were statistically significant (Student t-test, p<0.01). Parsimony analyses showed the rpoB gene evolves 4.8-fold faster than 16S rRNA. In addition, phylogeny of the rpoB gene separated each Anabaena strain more clearly compared with a 16S rRNA tree. These results suggest that the rpoB gene is a useful marker for the molecular phylogenetics and the species discrimination of Anabaena.

Preparation of Kimchi Containing Bifidobacterim animalis DY-64

  • Chae Myoung-Hee;Jhon Deok-Young
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.431-437
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    • 2006
  • Aero-tolerant microorganisms were isolated from healthy Koreans over the age of 95 years. The microorganisms were then identified based on their morphological and biochemical characteristics and 16S rDNA sequences. The growth properties of the isolated strains were investigated in kimchi. The characteristics of kimchi containing the microorganisms were studied microscopically, physicochemically, and organoleptically. Among 7 aero-tolerant strains, a strain with a 16S rDNA sequence exhibiting 99% homology with Bifidobacterim animalis strain B83 was selected and named B. animalis DY-64. The new strain showed a better acid resistance and salt resistance (p<0.05) than B. animalis ATCC 25527. After 15 days of fermentation in kimchi, the viability of B. animalis DY-64 was about 10%, and the kimchi had a better overall edible quality than conventional kimchi. Thus, it was found that the application of B. animalis DY-64 to kimchi preparation produced a good overall edible quality.

Isolation of a Marine Bacterium Capable of Biodegrading Poly(butylene succinate)

  • Lee, Sang Jun;Park, Eun Hee;Han, Yun Hee;Kim, Young Ok;Park, Seong Wook
    • Fisheries and Aquatic Sciences
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    • v.16 no.1
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    • pp.41-44
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    • 2013
  • We developed a poly(butylene succinate) (PBS) indicator plate and isolated a marine bacterial colony capable of biodegrading PBS based on the appearance of a clear zone. Growth of the PBS-2 isolate was observed over 4 days of culture at $37^{\circ}C$ in PBS-tryptone basal liquid medium, but not in PBS-deprived control medium. The PBS-2 isolate was named Paenibacillus sp. PBS-2 based on 16S rDNA gene sequencing. The PBS-biodegrading marine bacterium isolated in this study will contribute to the effective management of PBS waste problems in marine environments.